PT2501703E - Spiropiperidine compounds as orl-1 receptor antagonists - Google Patents

Abstract

An ORL-1 receptor antagonist of the formula: its uses, and methods for its preparation are described.

Description

ΡΕ2501703 1

DESCRIPTION

" SPIROPIPERIDINE COMPOUNDS AS RECEPTOR ANTAGONISTS ORL-1 "

Orphanin FQ (OFQ) / Nociceptin is a 17 amino acid peptide that has high affinity for the receptor coupled to the G protein ORL-1 (GPCR). The ORL-1 receptor is a Class A GPCR which is primarily expressed in the central nervous system and peripheral nervous system as well as in the gastrointestinal tract, smooth muscle, and immune system. Although structurally related to peptides / opioid receptors the OFQ / Nociceptin system does not exhibit significant cross-reactivity with classical peptides / opioid receptors and exhibits anti-opioid activity in vivo (e.g., ORQ / Nociceptin has been reported to exhibit anti-nociceptive properties).

Nociceptin / orphanin FQ (NOC / OFQ) receptor antagonists, specifically ORL-1 receptor antagonists have demonstrated anti-depressant activity and anoretic activity in numerous animal model studies for depression and eating behavior. As such, ORL-1 antagonists are considered useful in the treatment of depression and / or in the treatment of overweight, obesity, and / or weight maintenance after treatment for overweight or obesity. 2 ΡΕ2501703 WO 2003/095427 describes certain spiro-piperidinyl compounds as antagonists of ORL-1 for use as analgesics.

Yoshizumi, Takashi et al. (2008), Design, synthesis, and structure-activiy relationship study of a novel class of ORL-1 receptor antagonists based on N-biarylmethyl spiropiperidine, Bioorganic & Medicinal Chemistry Letters vol. 18, pp. 3778-3782, describes certain N-biarylmethyl-spiropiperidine compounds as selective ORL-1 antagonists. The present invention provides a family of 4 ', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] compounds with high potency antagonist for the ORL-1 receptor and high in vivo occupancy of the ORL-1 receptor in the CNS. In addition, some of the compounds have a favorable cardiotoxicological profile as determined through selectivity over hERG channel activity, as well as high selectivity over other physiologically important receptors (eg mu opioids, kappa and delta, serotonin, and dopamine). In addition, some of the compounds of the present invention have favorable biopharmaceutical and pharmacokinetic properties (e.g., solubility, oral exposure, and CNS permeability). Some of the compounds of the present invention exhibit reduced oxidant metabolism resulting in favorable oral bioavailability. Certain compounds have also demonstrated in animal models 3 ΡΕ2501703 that the compounds of the present invention are useful in the treatment of migraine. The present invention provides compounds of Formula I:

I wherein R1 is fluoro or chloro; R 2a and R 2b are each hydrogen or are each fluoro; R 3 is hydrogen, methyl, hydroxymethyl, or (C 1 -C 3) -alkoxymethyl; R 4 is selected from the group consisting of fluoro, chloro, cyano, cyanomethyl, (C 1 -C 3) alkyl, cyclopropyl, hydroxymethyl, methoxy, cyclopropylmethoxy, aminocarbonylmethoxy, (C 1 -C 3) alkoxy, cyclopropyloxymethyl, cyclopropylmethyl- 1-hydroxy-1-methylethyl, aminocarbonyloxymethyl, methylaminocarbonyloxymethyl, dimethylaminocarbonyloxymethyl, aminocarbonyl, aminocarbonylmethyl, -CH 2 -NR 5 R 6, hydroxyimine, methoxyimine, morpholin-4-yl, morpholin-4-ylmethyl, Ar 1 , 4 ΡΕ2501703 -CH2Ar1, tetrahydrofuran-2-yl, 3-oxomorpholin-4-ylmethyl, 2-oxopyrrolidin-1-ylmethyl, and 2-oxopiperidin-1-ylmethyl; R 5 is hydrogen, C 1 -C 3 alkyl, cyanomethyl, -C (O) -CH 3, or aminocarbonylmethyl; R6 is hydrogen or methyl; and

Ar1 is a moiety selected from the group consisting of imidizol-1-yl, imidizol-2-yl, 2-methylimidizol-1-yl, pyrazol-1-yl, 1,2,3-triazol- yl; 1,2,3-triazol-2-yl; 1,2,4-triazol-1-yl, isoxazol-3-yl, oxazol-5-yl, and 3-methyl-1,2,4-oxadiazol-5-yl; or a pharmaceutically acceptable salt thereof.

In another aspect of the invention there is provided a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, combined with a pharmaceutically acceptable carrier, diluent, or excipient. In one embodiment of this aspect of the invention, the pharmaceutical composition further comprises at least one additional therapeutic ingredient, such as an SSRI antidepressant such as, for example, fluoxetine. In addition, this aspect of the invention provides a pharmaceutical composition adapted for the treatment of depression comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, in combination with one or more of its pharmaceutically acceptable excipients, carriers, or diluents. In a further embodiment of this aspect of the invention there is provided a pharmaceutical composition adapted to the treatment of overweight, obesity and / or weight maintenance comprising a compound of Formula I or a pharmaceutically acceptable salt thereof in combination with a pharmaceutically acceptable salt thereof. or more of the pharmaceutically acceptable excipients, carriers, or diluents thereof. A further embodiment provides a pharmaceutical composition adapted to the treatment of migraine comprising a compound of Formula I or a pharmaceutically acceptable salt thereof in combination with one or more of the pharmaceutically acceptable excipients, carriers, or diluents thereof. The present invention further provides a method of treating depression in a mammal comprising administering to a mammal in need of such treatment an effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof. A further embodiment provides a method for treating depression in a mammal comprising administering to a mammal in need of such treatment an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof, and an effective amount of an anti- depressant SSRI, or a pharmaceutically acceptable salt thereof, such as fluoxetine. Other embodiments of the invention provide methods for treating overweight and / or obesity, and / or a method for weight maintenance comprising administering to a mammal in need of such treatment an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof. In a specific embodiment of these aspects of the invention, the mammal is a human.

This invention also provides a compound of Formula I or a pharmaceutically acceptable salt thereof for use in therapy. Within this aspect, the invention provides a compound of Formula I, or a pharmaceutically acceptable salt thereof, for use in the treatment of depression in mammals, particularly humans. The invention also provides a compound of Formula I, or a pharmaceutically acceptable salt thereof, in combination with an SSRI antidepressant, or a pharmaceutically acceptable salt thereof, such as, for example, fluoxetine, for use in the treatment of depression in mammals, particularly humans. In addition, this aspect of the invention includes any of the following: a compound of Formula I, or a pharmaceutically acceptable salt thereof, for use in the treatment of overweight; a compound of Formula I, or a pharmaceutically acceptable salt thereof, for use in the treatment of obesity; a compound of Formula I, or a pharmaceutically acceptable salt thereof, for use in maintaining weight (for weight maintenance), particularly after treatment of overweight or obesity; a compound of Formula I, or a pharmaceutically acceptable salt thereof, for use in the treatment of migraine.

Another aspect of this invention provides the use of the compound of Formula I, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of depression. Another embodiment of the invention provides the use of a compound of Formula I, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of overweight, obesity, and / or weight maintenance. Yet another embodiment of the invention provides the use of a compound of Formula I, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of migraine.

Compounds of this invention are bases, and accordingly react with various organic and inorganic acids to form pharmaceutically acceptable salts. Pharmaceutically acceptable salts of each of the compounds of the present invention are contemplated within the scope of the present application. The term " pharmaceutically acceptable salt " as used herein, refers to any salt of a compound of Formula I which is substantially non-toxic to living organisms. Such salts include those listed in the Journal of Pharmaceutical Science, 66, 2-19 (1977), which are known to those skilled in the art.

Abbreviations used herein are defined as follows: " BSA " means bovine serum albumin. ΡΕ2501703 " mCPP " means meta-chlorophenylpiperazine, a non-selective serotonin receptor agonist. " EDTA " means ethylenediamine tetraacetic acid. " EGTA " means ethylene glycol-tetra-acetic acid. " GTP " means guanosine triphosphate. " HEPES " means 4- (2-hydroxyethyl) -1-piperazine-ethanesulfonic acid. " HPLC " means high pressure liquid chromatography. " IC50 " means the concentration at which 50% of maximal inhibition is achieved. " LC / MS " means liquid chromatography followed by mass spectroscopy. " LC / MS / MS " means liquid chromatography followed by mass spectroscopy, followed by a second ionizing mass spectroscopy. " mFST " means mouse forced swimming test; an animal model for anti-depressant activity. " MS " means mass spectroscopy. " MS (ES +) " means mass spectroscopy using electrospray ionization. " NMR " means nuclear magnetic resonance. " RO Tracer " means 2 - [(2-Fluorophenyl) methyl] -3- (2-fluoro-spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) N, N-dimethylpropanamide. " RO " means occupation of the receiver. " SCX column " means strong cation exchange column. " SNAr " means nucleophilic aromatic substitution. " SSRI " means selective serotonin reuptake inhibitor. 9 ΡΕ2501703 " tBu " means a tertiary butyl moiety. " TLC " means thin layer chromatography. " XRD " means X-ray diffraction.

Preferred compounds of the present invention are compounds wherein: 1) R1 is chloro; 2) R 2a and R 2b are each fluoro; 3) R1 is chloro and R2a and R2b are each fluoro; 4) R1 is fluoro and R2a and R2b are each hydrogen; 5) R 3 is hydrogen, methyl, hydroxymethyl, or methoxymethyl; 6) R 3 is methyl; 7) R 3 is hydroxymethyl; 8) R1 is chloro, R2a and R2b are each fluoro, and R3 is methyl; 9) R1 is chlorine, R2a and R2b are each fluoro, and R3 is hydroxymethyl; 10) R 4 is fluoro, hydroxymethyl, methoxymethyl, or pyrazol-1-ylmethyl; 11) R 4 is fluoro; 12) R 4 is hydroxymethyl; 13) R 4 is methoxymethyl; 14) R4 is pyrazol-1-ylmethyl; 15) any of the preferred embodiments 1) to 9) wherein R 4 is fluoro; 16) any of the preferred embodiments 1) to 9) wherein R 4 is hydroxymethyl; 17) any of the preferred embodiments 1) to 9) wherein R 4 is methoxymethyl; 18) any of the preferred embodiments 1) to 9) wherein R 4 is pyrazol-1-ylmethyl;

Certain preferred compounds are [2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl ] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol; 3-methyl-1- [3- (pyrazol-1-ylmethyl) -2-pyridyl] pyrazol-4-yl] methyl] spiro [5H-thieno [ 2,3-c] pyran-7,4'-piperidine]; and [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1- (3- -fluoro-2-pyridyl) pyrazol-3-yl] methanol; or a pharmaceutically acceptable salt thereof, (i.e., Examples 1, 23, 53, 62, and 63)

A particularly preferred compound of the present invention is [2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1 yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol, or a pharmaceutically acceptable salt thereof, for example the L-tartrate salt and / or the HCl salt, such as exemplified in examples 1, 62 and 63.

Compounds in which R2a and R2b are each fluoro are preferred since the compounds have a more favorable pharmacokinetic profile, being more stable to 11 ΡΕ2501703 oxidizing metabolism. This has the general effect of improving the oral bioavailability of the compounds.

General chemistry

The compounds of the present invention may be prepared according to the following synthetic schemes by methods well known and considered in the art. Reaction conditions suitable for the steps of these schemes are well known in the art and suitable substitutions of solvents and co-reactants are within the skill of the art. Also, it will be appreciated by those skilled in the art that synthetic intermediates can be isolated and / or purified by various well-known techniques as required or desired, and that it is often possible to use several intermediates directly in the subsequent synthetic steps with little or no purification. In addition, those skilled in the art will appreciate that in some circumstances, the fact that the order in which the portions are introduced is not critical. The specific order of steps required to produce the compounds of Formula I depends on the specific compound to be synthesized, the starting compound, and the relative instability of the substituted moieties, as is well appreciated by the skilled chemists. All substituents, unless otherwise indicated, are as previously defined, and all reagents are well known and are considered in the art. ΡΕ2501703 12

Scheme 1

Using reductive amination reaction conditions, compound III is reacted with an appropriate substituted pyrazole-carbaldehyde and a reducing reagent such as sodium triacetoxyborohydride in a suitable solvent such as tetrahydrofuran at room temperature to afford the compound II. Under a suitable coupling condition, the compound II is coupled with the compound IV, wherein Hal is chloro, bromo, or iodo, with a suitable catalyst such as copper iodide, a suitable base such as potassium carbonate, in a solvent such as as toluene at elevated temperature to give compound I ', wherein R 4 is R 4 or a precursor of R 4. When Hal in compound IV is F or Cl, nucleophilic aromatic substitution (SNAr) is an alternative method for making compound I '. More specifically, compound II can 13? 2505070 react with compound IV in an appropriate solvent such as dimethylformamide with a suitable base such as potassium carbonate at elevated temperature to provide compound I '. The compound III may also be reacted with an appropriately substituted aldehyde compound V under reductive amination conditions as already mentioned to give the desired compound I '. When R 4 is a precursor of R 4, it is then converted to R 4 by known methods.

Scheme 2

xi x ix me

Compounds IIIa, IIIb, and IIIc may be obtained as shown in Scheme 2. Compounds XIV and XV are reacted in an appropriate solvent such as dichloromethane in the presence of a suitable acid such as trifluoroacetic acid. The resulting trifluoroacetate is basified with aqueous sodium hydroxide solution to give compound XIII as a free base. Compound XIII in a suitable solvent such as methyl t-butyl ether is treated with a solution of sulfuryl chloride in acetic acid at room temperature to give Compound VIII as a hydrochloride salt. Compound VIII is then protected with a nitrogen protecting group under conditions well known to one skilled in the art to give compound VII. (For example, see: Greene and Wuts, Protective Groups in Organic Synthesis, Third Edition, Chapters 2 and 7, John Wiley and Sons Inc., (1999)). Typically, the protecting group is a Boc (tert-Butyloxycarbonyl) group. Compound VII is reacted with N-bromosuccinimide in an appropriate solvent such as chlorobenzene with irradiation from a light source to give a bromide compound, which is then treated with a base solution such as aqueous sodium bicarbonate to obtain a hydroxyl compound. With or without isolation, the hydroxyl compound may be further oxidized under suitable oxidizing conditions such as potassium bromide, tetramethylpiperidine-17-oxide, and aqueous sodium hypochlorite solution to provide the desired ketone compound VI. Compound VI is then reacted with (bis (2-methylethyl) amino) sulfur trifluoride in an appropriate solvent such as tetrahydrofuran at elevated temperature, the product obtained is deprotected to provide compound IIIa. The compound of formula XIII is protected to give compound XII in a similar method to that used to make compound VII. Compound XII in a suitable solvent such as tetrahydrofuran at reduced temperature is treated with a suitable base such as lithium tetramethylpiperidine followed by addition of N-fluorobenzenesulfonimide to obtain a fluoride compound, which is deprotected with aqueous HCl 15 ΡΕ2501703 and basified with aqueous NaOH solution to provide compound IIIb.

By the same method as used to make said compound VI, compound IX can be obtained from compound XII through a three-step synthesis, such as bromination, hydroxylation, and oxidation. Each intermediate may be isolated as pure compound for additional reaction or subject to reaction without isolation as described in the synthesis of compound VI. Compound IX is then treated with a suitable halogen-metal exchange reagent such as butyllithium in a proprietary solvent such as tetrahydrofuran under reduced temperature, followed by a fluorination reagent such as N-fluoro-benzenesulfonimide to obtain the product fluorinated, which is then suitably deprotected to give the desired compound IIIc.

Scheme 3

Z = CMO. COOR R = Alkyl

Compounds of Formula V may be obtained such as 16 ΡΕ2501703 shown in Scheme 3. Compound XVI is reacted with compound XVII to obtain the compound of formula XV under SNAr or coupling conditions mentioned above for the conversion of compound II to compound I '. When Z is an ester group, it may be reduced to an alcohol first with a suitable reducing reagent such as lithium aluminum hydride in an appropriate solvent such as tetrahydrofuran. The alcohol is then oxidized to aldehyde with an appropriate oxidizing reagent such as manganese (IV) oxide in a solvent such as dichloromethane.

When R4 'is the precursor of R4, the transformation of R4' into R4 will include but is not limited to reactions such as reductive amination to provide a desired new amine; reducing an ester, ketone, or aldehyde to an alcohol, which may be further converted into an alkoxy compound or a carbamate; reduction of a nitrile to an amide or to an amine; the conversion of an ester into a heterocycle such as oxadiazole under appropriate condition. (For more examples, see: Richard C. Larock, Comprehensive Organic Transformations, Second Edition, Chapters 2 and 7, John Wiley and Sons Inc., (1999)).

The following Preparations and Examples are illustrative of methods useful for the synthesis of the compounds of the present invention. The names for many of the compounds illustrated in the preparations and examples are provided from structures drawn with 'Symyx Draw 3.1' or 'Autonom 2000 Name'. 17 ΡΕ2501703

Preparation 1: 4 ', 5'-Dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran]

To a solution of 17-tert-butoxycarbonyl-4-piperidone (185 g, 928.48 mmol) in dichloromethane (1300 mL) and stirred at 0 ° C was added 3-thiophene-ethanol (123.03 mL, 1.11 mol) room temperature. Then trifluoroacetic acid (280.82 mL, 3.71 moles) (5 min) is added dropwise while cooling with an ice / water bath (internal temperature = 14-30 ° C, caution: release of CO2) . The reaction mixture is gradually warmed to ambient temperature and stirred at that temperature for 20 h. The solvent is evaporated to give a crystalline solid beige after cooling in vacuo. The solid is slurried in methyl t-butyl ether (200 mL), filtered, washed with methyl t-butyl ether (2 x 1000 mL) and dried under vacuum to provide 4 ', 5'- hydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-one as a white solid in 95% yield. MS: (m / z): 210 (M + 1). 10M sodium hydroxide (220.36 mL, 2.20 mol) is added to a stirred suspension of 4 ', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-trifluoroacetate (285 g, 881.44 mmol) in dichloromethane (1 L) with cooling (ice / water bath) and the resulting mixture is stirred until a two phase mixture is obtained. The phases are separated and the aqueous layer is extracted with dichloromethane (2 x 200 mL). The combined organics are concentrated under vacuum to obtain a thick oil which is triturated with water to obtain a light yellow precipitate. The precipitate is filtered, washed with water (300 mL) and hexane (200 mL) and dried under vacuum at 35 ° C for 20 h to yield the title compound as a pale yellow solid in 86% yield. MS (m / z): 210 (M + 1).

Preparation 2: tert-Butyl spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-carboxylate

Spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] (60 g, 286.6 mmol) in 2-methyltetrahydrofuran (600 mL) is stirred at 22 ° C for 10 min. Then, tert-butoxycarbonyl and tert-butyl carbonate (65.6 g, 301 mmol) in 2-methyltetrahydrofuran (300 mL) is added dropwise. After 12 h, an aqueous solution of sodium chloride (250 mL) is added and the organic layer is separated. The aqueous layer is washed twice with 2-methyltetrahydrofuran (2 x 50 mL) and the organic layers are combined and washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound. with 99% yield. MS (m / z): 310 (M + 1). 19 ΡΕ2501703

Preparation 3: 2-Fluorospiro [4,5-dihydrothieno [2,2-c] pyran-7,4'-piperidine]

1. tert -Butyl 2,2,6,6-Tetramethylpiperidine 2-Fluorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-carboxylate ( 18.7 mL, 110.5 mmol) is added over tetrahydrofuran (200 mL), and the solution is cooled under nitrogen at -78 ° C. 2.5M solution of butyllithium in hexane (37.2 mL, 93 mmol) is added and the mixture is stirred for 30 min at -78 ° C. A solution of spiro [4,5-dihydrothieno [2,3-c] pyran-7,4 '-piperidine] -1'-methylpiperidine is added to a fresh solution of lithium-2,2,6,6-tetramethylpiperidine carboxylate (20 g, 58.2 mmol) in tetrahydrofuran (90 mL) maintaining the temperature below -70 ° C. After 20 min, a solution of N-fluorobenzenesulfonimide (30.26 g, 93.07 mmol) in tetrahydrofuran (200 mL) previously cooled under nitrogen at -20 ° C was added via cannula. After stirring for 1 h, water (20 mL) and aqueous ammonium chloride solution (50 mL) are added. The organic layer is then separated and the aqueous is washed twice with methyl t-butyl ether (2 x 25 mL). The organics are combined and the solvent is evaporated under reduced pressure. The crude material is purified by normal phase HPLC using hexane / methyl t-butyl ether as solvents to give 2-fluoro-spiro [4,5-dihydrothieno-20α] -2,1,1,1-d] [2,3-c] pyran-7,4 '-piperidine] -1'-carboxylate in 50% yield. MS (m / z): 328 (M + 1). 2. 2-Fluorospiro [4,5-dihydrothieno [2,2-c] pyran-7,4'-piperidine] 37% Hydrochloric acid (11.75 mL, 125.22 mmol) is added to a solution of Tert-Butyl 2'-fluoro-4 ', 5'-dihydro-1H-spiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-carboxylate (8.2 g, 25.04 mmol) in isopropyl alcohol (57.4 mL) at 45 ° C. The resulting solution is stirred at 45 ° C for 6.5 h. The solvent is concentrated to a yellow suspension. Water (50 mL) is added and the mixture is basified with 5N aqueous sodium hydroxide solution. The aqueous phase is extracted with ethyl acetate (3 x 100 mL) and the combined organic extracts are washed with brine (50 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give the title compound in 96% of yield. MS (m / z): 228 (M + 1).

Preparation 4: 2'-chloro-4 ', 5'-dihydroespiro [piperidine-4,7'-thieno [2,3-c] pyran hydrochloride]

A solution of 4 ', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] (50 g, 238.88 mmol) in a mixture of acetic acid (400 mL) and ether methyl-t-butyl ester (40 mL) is cooled to 15 ° C. Then a solution of sulfuryl chloride (21.20 mL, 262.77 mmol) in acetic acid (100 mL) is added dropwise over 40 min at that temperature (internal temperature = 15-22 ° C ) and the mixture is stirred at room temperature for 20 h. Then, a solution of sulfuryl chloride (11.56 mL, 143.33 mmol) in acetic acid (50 mL) is added dropwise at room temperature. The reaction mixture is stirred at room temperature for 30 min and then is added dropwise (30 min) over methyl t -butyl ether (1 L) by cooling with ice / water bath with stirring. A white suspension forms and the solid is filtered. To obtain a second crop of material, the filtrate is concentrated (refilling with methyl t-butyl ether via rotavap). The resulting solid is suspended in methyl t-butyl ether (300 mL), the suspension is stirred at reflux (bath: 100 ° C) and methanol (30 mL) is added until a cloudy suspension forms. The suspension is then cooled to room temperature overnight. The suspension is further cooled in an ice / water bath and filtered. The solid is washed with methyl t-butyl ether (50 mL) and combined with the first crop to give the title compound in 60% yield. MS (m / z): 244 (M + 1).

Preparation 5: 2'-Chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran]

22 ΡΕ2501703 2-Chlorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-carboxylate To a suspension of hydrochloric salt of 2- (140 g, 0.49 mol) in dichloromethane (1.12 L) is added triethylamine (67.25 g, 0.49 mol) in dichloromethane (1.12 L) (3.05 g, 0.025 mole) and di-t-butyl dicarbonate (111.22 g, 0.509 mole) were added in portions and the resulting mixture is stirred at room temperature overnight. The reaction is washed with 1 N HCl (2 x) and water. The organic phase is dried over magnesium sulfate, filtered and concentrated under reduced pressure to obtain 2-chlorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-carboxylate of tert-butyl in 53% yield. MS (m / z): 244 (M + 1-Boc). 2-tert-Butyl 4'-oxo-2'-chloro-4 ', 5'-dihydrospiro [piperidine-4,7' -thieno [2,3-c] pyran] -1-carboxylate

In a 5L coated reactor, N-bromosuccinimide (115.02 g, 639.77 mmol) is added to a solution of 2-chlorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4 ' -piperidine] -1'-carboxylate (200 g, 581.61 mmol) in chlorobenzene (1.60 L) at rt. The resulting suspension is irradiated 3X with 100W flask lamps placed almost in contact with the outer wall of the reactor and the reactor temperature is set at 45øC. After 4 h, N-bromosuccinimide (26.14 g, 145.40 mmol) is added and the temperature is 23 ΡΕ2501703 maintained at 40 ° C for 15 h. The reaction mixture is cooled to 0 ° C and methyl t -butyl ether (500 mL) is added. The solid is filtered and the solution is concentrated to about 1000 mL of chlorobenzene solution. Then methyl t-butyl ether (1000 mL) is added, the solids are filtered, and the filtrate is concentrated to obtain 600 mL of a chlorobenzene solution. Dimethyl sulfoxide (806.47 mL, 11.35 mol) and sodium bicarbonate (95.38 g, 1.14 mol) are added at room temperature. After stirring for 24 h at room temperature, water / ice (1000 mL) is added and the phases are separated. The organic phase is washed with water (2 x 1 L) and concentrated to give a solution in chlorobenzene. Then dichloromethane (1.2 L) is added and the mixture is cooled to 5øC (ice / water bath). Potassium bromide (20.27 g, 170.31 mmol) and 2,2,6,6-tetramethylpiperidine-N-oxide (4.43 g, 28.38 mmol) are added. Then a solution of 6% sodium hypochlorite in water (644.40 mL, 567.68 mmol) adjusted to pH = 9 with sodium bicarbonate (s) is added to the reaction mixture at 5 ° C and the resulting mixture is stirred for 1 h at 5øC to room temperature. Water (1 L) is added and the phases are separated. Organic phase is washed with water (2 x 0.5 L) and cooled with ice / water bath. Then Potassium Bromide (2.03 g, 17.03 mmol), 2,2,6,6-tetramethylpiperidine-α-oxide (0.05 g, 0.32 mmol) and a solution of sodium hypochlorite to 6% in water (128.88 mL, 113.54 mmol) adjusted to pH = 9 with solid sodium bicarbonate are added to the reaction mixture at 5 ° C and a resulting mixture is stirred for from 5 ° C to at ambient tem- perature. Then, water (1 L) is added and the phases are separated. The organic phase is washed with water (2 x 1 L) dried and concentrated to give a dark brown solid. The solid is triturated with hexane (500 mL), 5% methyl t-butyl ether / hexane (250 mL) and 10% methyl t-butyl ether / hexane (250 mL) to obtain 4'-oxo- Tert-butyl 2'-chloro-4 ', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-carboxylate as a light brown solid in 66 yield %. MS (m / z): 258 (M + 1 (Boc)). Tert-butyl 2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-carboxylate

To a 500 mL PFA flask charged with tetrahydrofuran (81 mL) is added (bis (2-methoxyethyl) amino) sulfur trifluoride (183.62 g, 829.94 mmol) and 4'-oxo-2'- chloro-4 ', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-carboxylate (135 g, 377.24 mmol). The resulting suspension is stirred at 70 ° C for 24h. It is then cooled to room temperature and slowly poured into a mixture of ice and saturated aqueous sodium bicarbonate solution (4 L) with stirring (gas evolution). Methyl t-butyl ether is used to transfer the remaining material from the flasks. After the gas evolution ceases, solid sodium bicarbonate is added with stirring until pH 8 is reached. The resulting mixture is extracted with methyl t-butyl ether (3 x 500 mL) until no product is detected by TLC in step 25 ΡΕ2501703 aqueous. The combined organics are washed with water (3 x 500 mL) and brine (500 mL), dried over sodium sulfate and concentrated to give a thick dark oil (250 g). The crude material is dissolved in dichloromethane and filtered through a plug of silica gel eluting with 10% methyl t-butyl ether / hexane (6 L) and methyl t-butyl ether / hexane (4 L). The fractions are collected until no product is detected by TLC (20% methyl t-butyl ether / hexane, Rf = 0.5). The filtrate is concentrated to give a light brown solid which is dried under vacuum at 40 ° C to constant weight to obtain 2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine- 4,7'-thieno [2,3-c] pyran] -1-carboxylate in 70% yield. MS (m / z): 324 (M + 1 tBu). 4'-Chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] hydrochloric acid (74% 12 mL, 789.78 mmol) is added to a solution of 2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3- c] pyran] -1-carboxylate (60 g, 157.96 mmol) in isopropyl alcohol (420 mL) at 45 ° C. The resulting solution is stirred at 45 ° C for 15 h. The mixture is then concentrated to 1/4 volume to give a white suspension. Water (100 mL) is added and the suspension is basified with 6N aqueous sodium hydroxide solution to obtain a two layer mixture which is extracted with methyl t-butyl ether (3 x 100 mL). The combined organic extracts are washed with brine (50 mL), dried over sodium sulfate, and concentrated to give a light brown solid which is dried under vacuum to constant weight to afford 97% title compound. MS (m / z): 280 (M + 1).

Preparation 6: 2,4,4-Trifluorospiro [5H-thieno [2,3-c] pyran-7,4'-piperidine]

H

F 1. tert-Butyl 2-bromo-4-hydroxy-spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-carboxylate N-bromosuccinimide (2 , 2 equiv) is added to a solution of tert-butyl spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-carboxylate (13.5 g) in chlorobenzene (108 mL). The resulting suspension is irradiated with 260 W balloon lamp overnight. Further, Bromosuccinimide (1.7 g) is added to the mixture and the mixture is irradiated with 260 W balloon lamp for 3 hours. The solvent is removed under reduced pressure affording a residue which is dissolved in acetone (650 mL) and a solution of silver nitrate (8.8 g) in water (650

mL) is added. The mixture is stirred at room temperature in the dark overnight. The mixture is filtered off and the acetone is evaporated. Ethyl acetate is added and the organic layer is washed with saturated aqueous sodium bicarbonate solution and brine. The organic layer is dried over sodium sulfate, filtered and the solvent is evaporated under reduced pressure. The residue is purified by standard Isco phase chromatography (eluent: 15-60% hexane / ethyl acetate) to obtain 2-bromo-4-hydroxy-spiro [4,5-dihydrothieno [2,3-c] ] pyran-7,4'-piperidine] -1'-carboxylate in 38% yield. 2-Bromo-4'-oxo-spiro [piperidine-4,7'-thieno [2,3-c] pyrimidin-4-yl] pyrazol-1-yl] -1-carboxylic acid tert-butyl ester

Potassium bromide (535.67 mg, 4.50 mmol) is added to a solution of 2-bromo-4-hydroxy-spiro [4,5-dihydrothieno [2,3-c] pyran-7,4 ' (7.28 g) and 2,2,6,6-tetramethylpiperidine-1-oxide (281.33 mg, 1.80 mmol) in dichloromethane (70 mL) at 0 ° C. In another vessel is added sodium bicarbonate in water 10% sodium hypochlorite (22.34 mL, 36.01 mmol) to pH 9. This sodium hypochlorite-sodium bicarbonate solution is added dropwise at 0 ° C and the resulting dark suspension is stirred at 0 ° C for 15 min. Dichloromethane (20 mL) and water (20 mL) are added and the phases are separated. The organic phase is washed with water (20 mL) and dried over sodium sulfate. The solvent is removed under reduced pressure to obtain tert-butyl 2'-bromo-4'-oxo-spiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-carboxylate in 99% Yield. 3-Bromo-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-4-yl] 7,4'-piperidine] -1'-carboxylic acid tert-butyl ester

In a 100 mL perfluoroalkoxy flask is added (bis (2-methoxyethyl) amino) sulfur trifluoride (5.16 mL, 27.96 mmol) to dry tetrahydrofuran (3.5 mL). Then tert-butyl 2'-bromo-4'-oxo-spiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-carboxylate (4.5 g, 11.19 mmol ). The solution is stirred at 70 ° C overnight. After this time, methyl t-butyl ether (30 mL) is added, and the reaction mixture is carefully poured into sodium bicarbonate (saturated aqueous solution) with cooling in an ice bath. CO2 liberation is observed and sodium bicarbonate (saturated aqueous solution) is added until pH 8. The mixture is extracted with methyl t-butyl ether. The organic layer is decanted, washed with brine (2x), dried over magnesium sulfate and the solvent evaporated under reduced pressure. The crude product is purified by normal Isco phase chromatography eluting with methyl t-butyl ether / hexane to give 3.2 g of 2-bromo-4,4-difluoro-spiro [5H-thieno [2,3-c] ] pyran-7,4'-piperidine] -1'-carboxylate. MS (m / z): 368/370 (M + 1-tBu / M + 3-tBu). 4,4,4,4-Trifluorospiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-carboxylate

To a solution of 2-bromo-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'- piperidine] -1'-carboxylate (1.99 g, 4.69 mmol) in tetrahydrofuran (50 mL) under nitrogen at -78 ° C. The mixture is stirred at -78 ° C for 1 hour and solid N -fluoro-benzenesulfonimide (3.69 g, 11.73 mmol) is added. The mixture is allowed to warm to room temperature and stirred at room temperature overnight. Saturated aqueous ammonium chloride solution is added and the organic phase is extracted with ethyl acetate, dried over sodium sulfate and the solvent is removed under reduced pressure. The crude material is purified by standard Isco-phase chromatography (3-12% hexane / ethyl acetate) to afford 1.3 g of 2,4,4-trifluoro-spiro [5H-thieno [2,3-c ] pyran-7,4'-piperidine] -1'-carboxylate which is further purified by HPLC to obtain 0.818 g of that compound. MS (m / z): 308 (M + 1-tBu). 5.4.4-Trifluoropyrrolo [5H-thieno [2,3-c] pyran-7,4'-piperidine] 2,4,4-Trifluorospiro [5H-thieno [2,3-c] pyran-7 , 4'-piperidine] -1'-carboxylate (2.00 g, 5.50 mmol) is dissolved in hydrochloric acid (4N in dioxane) (10 mL, 40 mmol). The mixture is stirred at room temperature for 1 h and then passed through a 50 g SCX cartridge (solid cation exchange) to give 1.3 g of the title compound after evaporation of the 2N ammonia fraction in methanol. MS (m / z): 264 (M + 1). 30 ΡΕ2501703

Preparation 7: 2-Chloro-4,4-difluoro-1 '- [(3-methyl-1H-pyrazol-4-yl) methyl] spiro [5H-thieno [2,3- c] pyran-7,4' -piperidine]

To a solution of 2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine (105 g, 375 mmol) in tetrahydrofuran (1.58 L ) is added 3-methyl-1H-pyrazole-4-carbaldehyde (43.40 g, 394.12 mmol) and the mixture is stirred at room temperature for 1 h. Then, powdered sodium triacetoxyborohydride (95.46 g, 450.42 mmol) is added in 3 portions. The mixture is stirred at room temperature for 15 h. Then, the reaction mixture is poured into a saturated aqueous solution of ice-sodium bicarbonate (400 mL). The phases are separated. The aqueous phase is extracted with ethyl acetate (100 mL). Combined organic layers are washed with 50% brine and a solid precipitates in the organic phase. The organic phase is concentrated to give 170 g of the title compound. MS (m / z): 374 (M + 1).

The compounds of Preparation 8-11 are prepared essentially as described in Preparation 7 using 1,2-dichloroethane as the solvent. 31 ΡΕ2501703

Prep. Chemical name Structure Physical data: N °. 2-Fluoro-1 '- [(3-methyl-1H-pyrazol-4-yl) methyl] spiro [4,5-dihydrothieno [2,3- L and 93 322 (MEL) c] pyran-7,4'-piperidine] 2-Chloro-1 '- [(3-methyl-1H-pyrazol-4-yl) -dihydrothieno [2,3-d] [3] benzazepine (M + 1) c] pyran-7,4'-piperidine] 2,4,4-Trifluoro-1 '- [(3-methyl-1H-10-pyrazol-4-yl) methyl] spiro [5H-thieno [2,3- 4'-piperidine] 2-Chloro-1 '- (1 H -pyrazol-4-ylmethyl) spiro [4,5-dihydrothieno [2,3- and 99- (4- (trifluoromethyl) -piperidine] fj

Preparation 12: 2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-theno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl ] -3-methyl-pyrazol-1-yl] pyridine-3-carbaldehyde

To a 250 mL flask is added copper (I) iodide (1.91 g, 10.03 mmol), 2-chloro-4,4-difluoro-1 '- [(3-methyl-1H-pyrazol-4 -yl] methyl] spiro [5H-thieno [2,3-c] pyrazine-7,4'-piperidine] (25 g, 66.87 mmol), potassium 32 ΡΕ2501703 carbonate (19.60 g, , 43 mmol), toluene (50 mL) and a stir bar. The reaction mixture is degassed through 5 vacuum / refill cycles. Then, 2-bromo-3-formyl-pyridine (18.66 g, 100.31 mmol) and trans-N, N'-dimethylcyclohexane-1,2-diamine (3.16 mL, 20.06 mmol). The reaction is stirred at room temperature for 5 minutes. It is then immersed in an oil bath preheated to 115øC and stirred for 15 h at that temperature. The reaction mixture is cooled to room temperature, diluted with 300 mL of ethyl acetate and filtered through celite. It is washed with ethyl acetate (100 mL) and aqueous ammonium hydroxide (3%) (4 x 100 mL) to remove traces of copper, and then washed with water (50 mL) and brine (50 mL) . The solution is dried over sodium sulfate. The solvent is evaporated in vacuo to give a tan solid. The solid was filtered through a pad of silica gel eluting with 2-propanol / dichloromethane (3% to 5% 2-propanol) to give the title compound in 69% yield. MS (m / z): 479 (M + 1).

Preparation 13: 2- [4 - [(2-Fluorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3- pyrazol-1-yl] pyridine-3-carbaldehyde

ΡΕ2501703 The compound of Preparation 13 is essentially prepared as described in Preparation 12 using 2-bromopyridine-3-carbaldehyde and 2-fluoro-1 '- [(3-methyl-1H-pyrazol-4-yl) methyl] spiro [ 4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine]. The residue is purified by normal Isco phase chromatography to give 83% of the title compound. MS (m / z): 427 (M + 1).

Preparation 14: N - [(2-Chloro-3-pyridyl) methyl] acetamide

To a solution of (2-chloro-3-pyridyl) methanamine (300 mg, 2.10 mmol) in dichloromethane (2 mL) at ambient temperature is added anhydrous acetic acid (198.88 æL, 2.10 mmol) and triethylamine (293.26 æl, 2.10 mmol). After 1 h the reaction is quenched with saturated aqueous sodium bicarbonate solution and extracted with dichloromethane. The organic layer was separated, dried over magnesium sulfate and the solvent evaporated in vacuo to give 184 mg of N - [(2-chloro-3-pyridyl) methyl] acetamide. MS (m / z): 185 (M + 1)

Preparation 15: (2-Bromo-3-pyridyl) methyl methanesulfonate

Br

To a 2 L round bottom flask (equipped with stir bar, under nitrogen and at 0 ° C) containing 2-bromo-3-pyridine-carboxaldehyde (22, 15 g, 119.08 mmol) and methanol (200 mL), sodium borohydride (1.35 g, 35.72 mmol) is added in three portions. After 1 h at 0 ° C water (200 mL) is added and the reaction is concentrated under reduced pressure to remove the methanol. The resulting residue is dissolved in ethyl acetate and washed with water (200 mL). The organic extract is washed with brine, dried over magnesium sulfate, filtered and concentrated to obtain 22 g of (2-bromo-3-pyridyl) methanol as a white solid. MS (m / z): 188 (M + 1), 190 (M + 3). 2. (2-Bromo-3-pyridyl) methyl methanesulfonate

Triethylamine (8.90 mL) is added to a round bottom flask containing a solution of (2-bromopyridin-3-yl) methanol (8 g, 42.55 mmol) in dichloromethane (212 mL) at 0 ° C, then is added methanesulfonyl chloride (3.95 mL) and the mixture is stirred at that temperature for 1.5 h. After this period the mixture is washed with water and the organic layer is decanted, dried over magnesium sulfate and the solvent is evaporated under reduced pressure to give 10.4 g of the title compound used without further purification. MS (m / z): 266, 268 (M + 1, M + 3). 35 ΡΕ2501703

Preparation 16: 2- (2-Bromo-3-pyridyl) propan-2-ol

Diisopropylamide (2M, 12.5 mL, 25 mmol) is dissolved in tetrahydrofuran (40 mL) and cooled to -78 ° C. 2-Bromopyridine (3.9 g, 25 mmol) is added dropwise and the reaction is stirred for 3 h before adding acetone (1 mL, dried over freshly activated molecular sieves) and allowing to warm to room temperature. The reaction is quenched with saturated ammonium chloride and extracted with ethyl acetate. The organic layer is dried over magnesium sulfate and evaporated. The residue is purified by flash column chromatography on silica, eluting with 20% hexane / methyl tert-butyl ether to give the title compound (1.4 g, 26%). 1H NMR (d6 -dimethylsulfoxide): 8.23 (dd, J = 1.9, 4.5 Hz, 1H), 8.19 (dd, J = 2.0, 7.8 Hz, 1H) , 7.44 (dd, J = 4.5, 7.7 Hz, 1H), 5.43 (s, 1H), 2.12 (s, 1H), 1.46 (s, 6H).

Preparation 17 Ethyl 3-methyl-1H-pyrazole-4-carboxylate

Sulfuric acid (18 mL, 337.69 mmol) is added to a suspension of 1H-pyrazole-4-carboxylic acid, 3-methyl- (10 g, 79.29 mmol) in ethanol (90 mL) and the mixture is stirred at 85 ° C for 20 h. After this time, the solvent is partially removed. The residue is basified with 36 ΡΕ2501703 solution of 1M sodium hydroxide to pH 6-7 and extracted with dichloromethane. The organic layer is decanted, dried over magnesium sulfate and the solvent evaporated under reduced pressure to give 10.3 g of the title compound which is used without further purification. MS (m / z): 155 (M + 1).

Preparation 18: Methyl 2-fluoropyridine-3-carboxylate

The 2 M lithium diisopropylamide (11.3 mL, 22.5 mmol) is slowly added to a solution of 2-fluoropyridine (2 g, 20.5 mmol) in tetrahydrofuran (90 mL) under nitrogen atmosphere at -78Â ° C. After 4 h at that temperature methyl chloroformate (1.9 mL, 24.6 mmol) is added and the mixture is stirred for an additional hour at -78 ° C and allowed to reach room temperature overnight. The reaction mixture is slowly poured into water and extracted into diethyl ether. The organic layer is washed with brine, dried over sodium sulfate and the solvent is evaporated in vacuo. The residue is purified by standard Isco phase chromatography eluting with hexane / diethyl ether (9/1) to give 495.2 mg of the title compound. MS (m / z): 156 (M + 1).

Preparation 19: 1- (3-Fluoro-2-pyridyl) -3-methyl-pyrazole-4-carbaldehyde 37 ΡΕ2501703 Potassium t-butyl alcohol (29.25 g, 260.68 mmol) is added to a solution of 3-methyl-1H-pyrazole-4-carbaldehyde (26.00 g, 236.12 mmol) in dimethylformamide (250 mL) cooled with an aqueous bath. The mixture is stirred at room temperature for 10 min. Then, 2,3-difluoropyridine (25 g, 217.24 mmol) is added and the mixture is stirred at room temperature for 20 h. The mixture is poured into an ice / water mixture and extracted into ethyl acetate (3 x 20 mL). Organic are dried over magnesium sulfate and solvent evaporated in vacuo. The residue is purified by silica gel chromatography using hexane / isopropyl alcohol as the eluent to obtain a crude material with residual dimethylformamide. The material is then dissolved in ethyl acetate (100 mL) and washed with water (3 x 20 mL). The aqueous phase is extracted with ethyl acetate (2 x 20 mL). The combined organic extracts are dried over sodium sulfate and concentrated to give 19 g of the title compound. MS (m / z): 206 (M + 1).

Preparation 20: 2- (4-Formyl-3-methyl-pyrazol-1-yl) pyridine-3-carbonitrile

The compound of Preparation 20 is essentially 38æE2501703 prepared as described in Preparation 19 using 3-methyl-1H-pyrazole-4-carbaldehyde and 2-fluoropyridine-3-carbonitrile. The title compound is obtained in 23% yield. MS (m / z): 213 (M + 1).

Preparation 21: methyl 2- (4-formyl-3-methyl-pyrazol-1-yl) pyridine-3-carboxylate

A mixture of 3-methyl-1H-pyrazole-4-carbaldehyde (300 mg, 2.72 mmol), potassium carbonate (565 mg, 4.08 mmol), methyl 2-fluoropyridine-3-carboxylate (507 mg, 3.27 mmol) and dimethylformamide (2 mL) is stirred at 60 ° C for 16 h. Water is added and the compound is extracted into ethyl acetate. The organic layer is washed with brine, dried over sodium sulfate and the solvent is evaporated. The crude mixture is purified by normal Isco phase chromatography using hexane: ethyl acetate, 2: 1 as eluant to afford 422 mg of the title compound. MS (m / z): 246 (M + 1). The compound of Preparations 22-23 can be prepared essentially as described in Preparation 21 using the corresponding 2-chloropyridyl reagent. 39 ΡΕ2501703

Preparation 24: 1- [3- (Dimethylaminomethyl) -2-pyridyl] -3-methyl-pyrazole-4-carbaldehyde

1. Ethyl 1- [3- (dimethylaminomethyl) -2-pyridyl] -3-methyl-pyrazole-4-carboxylate

This compound is prepared essentially as described in Preparation 7 using ethyl 1- (3-formyl-2-pyridyl) -3-methylpyrazole-4-carboxylate and dimethylamine in 95% yield and is used without further purification after processing . [1- [3- (Dimethylaminomethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methanol To a solution of 1- [3- (217.7 g, 0.75 mmol) in tetrahydrofuran (5 mL) cooled to 0 ° C under nitrogen , 1M lithium aluminum hydride in tetrahydrofuran (0.9 mL, 0.9 mmol) is added and the mixture is stirred at that temperature for 5 minutes. The reaction is quenched with saturated aqueous ammonium chloride solution and the crude product is extracted with ethyl acetate. The organic layer is separated, dried over sodium sulfate and the solvent is evaporated in vacuo. In this particular example the compound is not isolated in the organic extraction and is finally separated from the aqueous solution with an SCX cartridge to obtain 146 mg (78%) of [1- [3- (dimethylaminomethyl) -2-pyridyl] -3-methyl- pyrazol-4-yl] methanol which is used without further purification. MS (m / z): 247 (M + 1). 3. 1- [3- (Dimethylaminomethyl) -2-pyridyl] -3-methyl-pyrazole-4-carbaldehyde 3,3,3-Triacetoxy-3-iodophthalide (0.553 g, 1.3 mmol) is added to a solution of [1- [3- (dimethylaminomethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methanol (0.146 g, 0.59 mmol) in dichloromethane (4 mL) at room temperature. After one hour the reaction is quenched by addition of 2N sodium carbonate solution and the compound is extracted into dichloromethane. The organic layer is separated, dried over magnesium sulfate and the solvent evaporated in vacuo to give 126 mg of 1- [3- (dimethylaminomethyl) -2-pyridyl] -3-methyl-pyrazole-4-carbaldehyde which is used without further purification. MS (m / z): 245 (M + 1). 41 ΡΕ2501703

Preparation 25: 1- (3-Fluoro-2-pyridyl) -3- (isopropoxymethyl) pyrazole-4-carbaldehyde

To a solution of ethyl 1- (3-fluoro-2-pyridyl) -3-methyl-pyrazole-4-carboxylate 4-carboxylate (10.2 g, 40.9 mmol) in chlorobenzene (205 mL) is added N-bromosuccinimide (8 g) at room temperature. The reaction mixture is stirred and irradiated under a 100 W lamp at room temperature overnight. Further N-bromosuccinimide (2 g) is then added and after 2 h, no starting material is detected. The solids are filtered and the solvent is evaporated. The crude product is purified by normal Isco phase chromatography using dichloromethane and methanol as eluant to give 5.3 g of ethyl 3- (bromomethyl) -1- (3-fluoro-2-pyridyl) pyrazole-4-carboxylate. MS (m / z): 328; 330 (M + 1; M + 3). 2. Isopropyl 1- (3-fluoro-2-pyridyl) -3- (isopropoxymethyl) pyrazole-4-carboxylate To a solution of 3- (bromomethyl) -1- (3-fluoro-2-pyridyl) pyrazole-4 carboxylate (1.5 g, 4.57 mmol), 42 ΡΕ2501703 in propan-2-ol (23 mL), lithium isopropoxide (3.02 g) is added and the mixture is stirred overnight. another at room temperature. After that time additional lithium isoproxide (4.5 g) is added in three 1.5 g portions each at 16 hours. After the last addition, the reaction mixture is finally stirred at 40 ° C for 16 hours. The solvent is evaporated and the residue is dissolved in ethyl acetate and washed with brine. The organic layer is decanted and dried over sodium sulfate. The solvent is evaporated and the residue is purified by normal Isco-phase chromatography using dichloromethane and methanol as eluant to give 317 mg of 1- (3-fluoro-2-pyridyl) -3- (isopropoxymethyl) pyrazol-4- carboxylate. MS (m / z): 322 (M + 1). 3. [1- (3-Fluoro-2-pyridyl) -3- (isopropoxymethyl) pyrazol-4-yl] methanol To a solution of 1- (3-fluoro-2-pyridyl) -3- (isopropoxymethyl) pyrazole- 4-carboxylate (316 mg, 0.98 mmol) in diethyl ether (4.9 mL) under nitrogen and cooled to 0 ° C is added 1M lithium aluminum hydride in tetrahydrofuran (1.2 mL ). The mixture is stirred at 0 ° C for 1 hour. Water (46 æL) is added and stirred for 5 min, then 15% aqueous NaOH (46 æL) and water (138 æL) are successively added. The solids are filtered and the solvent is evaporated to obtain 241 mg of [1- (3-fluoro-2-pyridyl) -3- (isopropoxymethyl) pyrazol-4-yl] methanol which is used without further purification. MS (m / z): 266 (M + 1). 43 ΡΕ2501703 4. 1- (3-Fluoro-2-pyridyl) -3- [isopropoxymethyl] pyrazole-4-carbaldehyde

A mixture of [1- (3-fluoro-2-pyridyl) -3- (isopropoxymethyl) pyrazol-4-yl] methanol (241 mg, 0.91 mmol) and manganese (IV) oxide (791 mg) is stirred in dichloromethane (5 mL) at room temperature overnight. After this period, the reaction mixture is filtered over celite and the solvent evaporated to obtain 197 mg of the title compound which is used without further purification. MS (m / z): 264 (M + 1).

Preparation 26: 1- (3-Fluoro-2-pyridyl) -3- (methoxymethyl) pyrazole-4-carbaldehyde

1. Ethyl 1- (3-fluoro-2-pyridyl) -3- (methoxymethyl) pyrazole-4-carboxylate To a solution of 3-bromomethyl-1- (3-fluoro-pyridin- 2-yl) -1H-pyrazole-4-carboxylic acid (1.13 g, 3.44 mmol) in acetone (23 mL), potassium iodide (200.1 mg), potassium carbonate (865, 33 mg) and methanol (4.18 mL). The mixture is stirred at room temperature for 48 hours. Methyl ester and ethyl ester of the desired compound are detected. The solids are evaporated. The residue is diluted with dichloromethane and washed with water. The organic layer is decanted, dried over magnesium sulfate and the solvent evaporated. The crude product is purified by standard Isco-phase chromatography eluting with ethyl acetate and hexane to afford 599 mg of a mixture of methyl and ethyl esters. MS (m / z): 266 (M + 1) and 280 (M + 1). 2. [1- (3-Fluoro-2-pyridyl) -3- (methoxymethyl) pyrazol-4-yl] methanol

This compound is essentially prepared as described in step 3 of preparation 25 (reduction step) in 93% yield and using as starting material 1- (3-fluoro-2-pyridyl) -3- (methoxymethyl) pyrazole- 4-carboxylate (in a mixture also containing the methyl ester). [1- (3-Fluoro-2-pyridyl) -3- (methoxymethyl) pyrazol-4-yl] -methanol is used without further purification. MS (m / z): 238 (M + 1). 3. 1- (3-Fluoro-2-pyridyl) -3- (methoxymethyl) pyrazole-4-carbaldehyde

This compound is essentially prepared as described in step 4 of preparation 25 (oxidation step) in 91% yield and using as starting material [1- (3-fluoro-pyridin-2-yl) -3-methoxymethyl- 1H-pyrazol-4-yl] -methanol. The title compound is used without further purification. MS (m / z): 236 (M + 1). 45 ΡΕ2501703

Preparation 27: 3-Methyl-1- (3-morpholino-2-pyridyl) pyrazole-4-carbaldehyde or

A mixture of 1- (3-fluoro-2-pyridyl) -3-methyl-pyrazole-4-carbaldehyde (230 mg, 1.12 mmol) and morpholine (2 mL, 22.93 mmol) is heated under micro- (160 ° C temperature, ramp time: 2 min, holding time: 10 min, power: 250 W). The excess morpholine is removed in vacuo and the residue is diluted in dichloromethane and washed with saturated aqueous sodium bicarbonate solution. The organic layer is separated, dried over magnesium sulfate and the solvent evaporated in vacuo. The residue is purified by normal Isco phase chromatography using hexane / ethyl acetate as eluent (50 to 100% in ethyl acetate) to give 72% of the title compound. MS (m / z): 273 (M + 1).

Preparation 28: 2-Bromo-3- (methoxymethyl) pyridine

To a solution of (2-bromopyridin-3-yl) methanol (20 g, 106.37 mmol) in 140 mL of tetrahydrofuran is added 60% sodium hydride in mineral oil (6.38 g, 46 ΡΕ2501703 , 5 mmol) at 0 ° C under a nitrogen atmosphere. The solution is stirred at 0 ° C for 30 minutes. Methyl iodide (7.95 mL, 127.6 mmol) is added over the solution and the mixture is stirred at room temperature overnight. The mixture is quenched by adding water and the crude product is extracted with ethyl acetate. The organic layer is separated, dried over magnesium sulfate and the solvent evaporated in vacuo to give 2-bromo-3- (methoxymethyl) pyridine in 98% yield which is used without further purification. MS (m / z): 202 (M + 1), 204 (M + 3).

The compounds of Preparation 29-30 are prepared essentially as described in Preparation 28 using the corresponding alkyl iodide reagents.

Preparation 31: 2-Bromo-3- (isopropoxymethyl) pyridine 47 ΡΕ2501703 1. 2-Bromo-3- (bromomethyl) pyridine

A 25 mL Schlenk flask is charged with 2-bromo-3-pyridine benzyl alcohol (1 g, 5.32 mmol) and dichloromethane (15 mL). The resulting solution was cooled to 0 ° C in a vigorously stirred magnetic salt-water / ice bath. Phosphorus tribromide (0.55 mL, 5.80 mmol) is then added dropwise to control the exotherm to below 10øC. A thick sludge resulted and after addition, the bath is removed and the sludge is heated to room temperature. The mixture is then gradually heated to reflux for 3 h and then to room temperature overnight.

About the pre-cooled mixture at 0 ° C, ice-water is added and then pH adjusted to basic by adding 2M sodium carbonate solution. The organic layer is separated and the aqueous solution is washed twice with dichloromethane (2 x 10 mL). All the organics are combined, dried over magnesium sulfate, filtered and the solvent evaporated under reduced pressure to afford 1.2 g of 2-bromo-3-bromomethyl-pyridine which is used without further purification. 2-Bromo-3- (isopropoxymethyl) pyridine To a solution of isopropyl alcohol (7.31 mL, 95.65 mmol) in dimethylformamide (50 mL) at 0 ° C is added sodium hydride (3.63 g , 90.87 mmol), and the mixture is stirred at 48 ΡΕ2501703 at room temperature for 30 minutes (this reaction is performed using a shield as a special shield). Then, 2-bromo-3-bromomethyl-pyridine (6 g, 23.91 mmol) in 10 mL of dimethylformamide is added via syringe and the mixture is stirred at room temperature for 1 h. Water is added and the mixture is extracted with ethyl acetate, washed with brine and water, dried over magnesium sulfate, filtered and the solvent evaporated under reduced pressure. The crude material is purified by Isco normal phase chromatography using ethyl acetate and hexane as eluent (5 to 25% ethyl acetate gradient) to give 3.67 g of the title compound. MS (m / z): 230.232 (M + 1, M + 3)

Preparation 32: 2- (2-Bromo-3-pyridyl) acetonitrile

Br

A mixture of 2-bromo-3-bromomethylpyridine (840 mg, 3.35 mmol) and sodium cyanide (187.99 mg, 3.68 mmol) is stirred and refluxed for 1 h in a mixture of water ( 10 mL) -ethanol (2 mL). The reaction mixture is diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate solution. The organic layer is separated, dried over magnesium sulfate and the solvent evaporated in vacuo to give 83% of the title compound which is used without further purification. MS (m / z): 197 (M + 1), 199 (M + 3). 49 ΡΕ2501703

Preparation 33: 1- [3- (Cyclopropoxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carbaldehyde

1. Ethyl 1- [3- (hydroxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carboxylate To a solution of 1- (3-formyl-pyridin-2-yl) - 3-methyl-1H-pyrazole-4-carboxylic acid (1 g, 3.9 mmol) in methanol (25 mL) is added sodium borohydrate (220.2 mg, 5.8 mmol). The mixture is stirred at room temperature for 2 h. The solvent is removed and the crude product is dissolved in ethyl acetate and washed with saturated aqueous ammonium chloride solution. The organic layer is separated, dried over sodium sulfate, filtered and the solvent evaporated under reduced pressure to give 1 g of ethyl 1- [3- (hydroxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carboxylate . MS (m / z): 262 (M + 1) 2. Ethyl 3-methyl-1- [3- (vinyloxymethyl) -2-pyridyl] pyrazole-4-carboxylate

(5.1 mg, 15.3 mmol) and 4,7-diphenyl-phenanthroline (5.1 mg, 15.3 pmoles) are dissolved in n-butyl vinyl ether (6.1 mL, 61.2 mmol) and 1- (3-hydroxymethyl-pyridin-4-yl) -2-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (800 mg, 3.1 mmol) and added a few drops of triethylamine. The flask is sealed and shaken at 75 ° C for 24 hours. The mixture is then cooled to 24 ° C and filtered through a pad of celite. The solvent is removed under reduced pressure and the residue is purified by standard Isco-phase chromatography using hexane and ethyl acetate as eluents (5-20% in ethyl acetate) to give 605 mg of 3-methyl-1- [3 - (vinyloxymethyl) -2-pyridyl] pyrazole-4-carboxylate. MS (m / z): 288 (M + 1) 3. Ethyl 1- [3- (cyclopropoxymethyl) -2-pyridyl] -3-methylpyrazole-4-carboxylate

Diethylzinc 1.0 M in hexane (365.7 æl, 3.5 mmol) is added under nitrogen to dichloromethane (0.9 mL). The solution is cooled to 0øC and a solution of trifluoroacetic acid (268.4 æl, 3.5 mmol) in dichloromethane (0.9 mL) is added very slowly. After 20 min a solution of diiodomethane (286.1 μL, 3.5 mmol) in dichloromethane (0.9 mL) is added. After a further 20 min, a solution of 3-methyl-1- (3-vinyloxymethyl-pyridin-2-yl) -1H-pyrazole-4-carboxylic acid ethyl ester (510 mg, 1.8 mmol) in dichloromethane (0.9 mL). The mixture is stirred at room temperature overnight. 5% HCl is added and the aqueous layer is extracted with ethyl acetate. The organic layer is separated, dried over sodium sulfate and the solvent is removed under reduced pressure. The crude product is purified by standard Isco 51 ΡΕ2501703 phase chromatography using hexane and ethyl acetate as eluents (5-20% in ethyl acetate) to obtain 310 mg of 1- [3- (cyclopropoxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carboxylate. MS (m / z): 302 (M + 1) 4. 1- [3- (Cyclopropoxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carbaldehyde To a solution of 1- ( 2-yl) -3-methyl-1H-pyrazole-4-carboxylic acid (410 mg, 1.03 mmol) in tetrahydrofuran (0.15 M) at 0øC under nitrogen, is added 1.0M lithium aluminum hydride in tetrahydrofuran (1.2 equiv). The mixture is stirred at room temperature for 1 h and saturated aqueous ammonium chloride solution is added and the aqueous layer is extracted with ethyl acetate. The organic layer is separated, dried over sodium sulfate and the solvent is removed under reduced pressure. The crude material is dissolved in dichloromethane (14 mL) and manganese (IV) oxide (1.2 g, 13.9 mmol) is added. The mixture is stirred at room temperature overnight and then filtered through a pad of celite and the solvent is evaporated under reduced pressure. The crude product is passed through a pad of silica gel (eluent: ethyl acetate and dichloromethane). The solvent is removed under reduced pressure to give 90 mg of the title product. MS (m / z): 258 (M + 1). 52 ΡΕ2501703

Preparation 34: 1- [3- (Methoxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carbaldehyde

O

1. Ethyl 1- [3- (methoxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carboxylate

This compound is prepared essentially as described in Preparation 12 using ethyl 3-methyl-1H-pyrazole-4-carboxylate and 2-bromo-3- (methoxymethyl) pyridine in 83% yield. MS (m / z): 276 (M + 1). 2. [1- [3- (Methoxymethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methanol

This compound is prepared essentially as described in Preparation 24 (step 2) using ethyl 1- [3- (methoxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carboxylate in 88% yield. MS (m / z): 234 (M + 1). 3. 1- [3- (Methoxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carbaldehyde

A mixture of [1- [3- (methoxymethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methanol (2.2 g, 9.4 mmol) and manganese (IV) oxide (19 g , 188.8 mmol) is stirred in dichloromethane 53 ΡΕ2501703 (20 mL) at room temperature overnight. The reaction mixture is filtered over celite and the solvent evaporated under reduced pressure to give 1.8 g of the title compound which is used without further purification. MS (m / z): 232 (M + 1).

Preparation 35: 2-Bromo-3- (imidazol-1-ylmethyl) pyridine

To a solution of (2-bromo-3-pyridyl) methyl methanesulfonate (2.5 g, 9.39 mmol) in ethanol (46.97 mL) at room temperature, potassium carbonate (2.62 g) and 1 H -imidazole (1.31 g). The mixture is stirred at 50 ° C. A thick emulsion is formed and 10 ml of ethanol is added and the emulsion species is stirred at 50 ° C overnight. Water is added until all solids are dissolved. Then, it is concentrated under reduced pressure and the residue is diluted with ethyl acetate and washed with sodium bicarbonate (saturated aqueous solution). The organic layer is decanted, dried over magnesium sulfate and the solvent evaporated in vacuo. The crude material is purified by standard Isco phase chromatography using dichloromethane and methanol (98/2 to 90/10) as eluent to give 1.42 g of the title compound. MS (m / z): 238.240 (M + 1, M + 3). 54 ΡΕ2501703

The compounds of Preparations 36-40 are prepared essentially as described in Preparation 35 from the corresponding nitrogen hetero rings.

Prep. No. Chemical name Structure Yield (%) Physical data: MS (m / z) 36 2-Bromo-3- (1,2,4-triazol-1-ylmethyl) pyridine N (V 39 239, 241 (MH + M + 3) 37 2-Bromo-3- (triazol-1-ylmethyl) pyridine / Î'and 33a 239, 241 (MH, IYH-3) 2-Bromo-3 - [(2-methylimidazol-1-yl) -piperidin-2-yl] yl) methyl] pyridine hydrochloride (decomp.), 252, 254 (MH, M + 3) a) The compounds are obtained in the same reaction.

Preparation 41: 2 - [(2-Bromo-3-pyridyl) oxy] acetamide

Br

To a solution of 2-bromo-3-pyridinol (1.5 g, 8.49 mmol) in tetrahydrofuran (15 mL) at 0 ° C is added 55 Ρ TE2501703 60% sodium hydride in mineral oil (509, 44 mg, 12.74 mmol) and the mixture is stirred at that temperature for 30 min. Acetamide, 2-chloro- (16.98 mmol) is then added and the mixture is stirred at room temperature over the weekend. The mixture is quenched with water and extracted with ethyl acetate. The organic layer is separated dry over magnesium sulfate and the solvent evaporated in vacuo to give the title compound in 75% yield. The compound is used without further purification. MS (m / z): 231 (M + 1), 233 (M + 3).

Preparation 42: 2-Bromo-3- (cyclopropylmethoxy) pyridine

To a solution of 2-bromo-3-pyridinol (500 mg, 2.83 mmol) in dimethylformamide (3 mL) is added 60% sodium hydride in mineral oil (169.81 mg, 4.25 mmol ). The mixture is stirred at that temperature for 15 min and bromomethylcyclopropane (329.66 æL, 3.40 mmol) is added. Then the reaction is stirred at room temperature for 4 hours and quenched with ethyl acetate. The organic layer is separated, washed with brine, dried over magnesium sulfate and the solvent evaporated in vacuo. The crude product is passed through a short silica gel plug and the compound eluted with dichloromethane to give 23% of the titled compound. MS (m / z): 228 (M + 1), 230 (M + 3). 56 ΡΕ2501703

Preparation 43: Ethyl 1- (3-fluoro-2-pyridyl) -4-formyl-pyrazole-3-carboxylate

1. Ethyl 4-formyl-1H-pyrazole-3-carboxylate

A 100 mL round bottom flask is charged with semicarbazide hydrochloride (2 g, 17.93 mmol), ethyl pyruvate (11.8 mL) and sodium ethanoate (2.97 g). Then water (9 mL) is added. The mixture is stirred at room temperature for 90 minutes, then filtered and the solids washed with more water. The white solid is dried under vacuum in an oven (40 ° C) overnight. 2.66 g of pure ethyl 2- (carbamoylhydrazono) propanoate are obtained. Phosphoryl chloride (3.28 mL) is added dropwise to dimethylformamide (7.13 mL) at 0 ° C. The cooling bath is removed to reach room temperature. The mixture is then heated to 40 ° C and ethyl 2- (carbamoylhydrazono) propanoate previously obtained (2.66 g, 15.36 mmol) is added in several portions over 15-20 min (gas evolution during the addition). When the internal temperature is 55øC, the heating bath is removed so that the internal temperature is maintained in the range of 60-70øC. The mixture is then heated to 80 ° C. 57 ΡΕ2501703

Almost no further gas release is observed once the reaction is stabilized at 80 ° C. The resulting suspension is stirred at 80 ° C for 2 h. The reaction mixture is poured into ice / water (30 mL), and stirred to give a suspension. 50% sodium hydroxide solution is added dropwise until it reaches pH 10. The solution is stirred at 50 ° C for 5 min. The solution is then cooled with an ice / water bath and 35% aqueous hydrochloric acid solution is added until it reaches pH 7. The mixture is extracted with ethyl acetate. The organic layer is decanted, dried over magnesium sulfate and the solvent is evaporated under reduced pressure. The residue is triturated with hexane and a few drops of dichloromethane to give 1.6 g of ethyl 4-formyl-1H-pyrazole-3-carboxylate which is used in the next step without further purification. MS (m / z): 169 (M + 1). 2 . Ethyl 1- (3-fluoro-2-pyridyl) -4-formyl-pyrazole-3-carboxylate

A mixture of ethyl 4-formyl-1H-pyrazole-3-carboxylate (1.6 g, 9.52 mmol), 2,3-difluoropyridine (1.20 g) and potassium carbonate (1.97 g) in dimethylformamide (8 mL) is heated and stirred at 100 ° C for 1.5 h. Then, the reaction mixture is stirred at room temperature overnight. Water is added, and the organic phase is extracted with ethyl acetate, decanted, dried over magnesium sulfate and the solvent evaporated under reduced pressure. The crude product is purified by normal phase chromatography 58 ΡΕ2501703

Bait using ethyl acetate and hexane to give 711 mg of the title compound. MS (m / z): 264 (M + 1).

Preparation 44: 3-Methyl-1- (3- (tetrahydrofuran-2-yl) pyridin-2-yl) -1H-pyrazole-4-carbaldehyde

1. 3- (2,5-Dihydrofuran-2-yl) -2-fluoropyridine To a screw cap test tube containing potassium acetate (440 mg, 4.5 mmol), tetrabutylammonium bromide (1 , 4 g, 4.5 mmol) and 4A molecular sieves in dry dimethylformamide (1.8 mL) are added under nitrogen 2-fluoro-3-iodopyridine (400 mg, 1.8 mmol), 2,3-dihydro fura (1.4 mL, 18 mmol) and palladium acetate (20 mg, 0.09 mmol). The reaction tube is quickly sealed and shaken overnight at room temperature. The mixture is diluted with diethyl ether and the filtrate is filtered over celite. the filtrate is washed with water, dried over magnesium sulfate, filtered and the solvent evaporated in vacuo. The resulting residue is purified by silica gel using Isco normal phase chromatography eluting with hexane: acetone (5 to 30% gradient in acetone) to afford 170 mg of 3- (2,5-dihydrofuran-2-yl ) -2-fluoropyridine. MS (m / z): 166 (M + 1). 59 ΡΕ2501703 2. 2-Fluoro-3- (tetrahydrofuran-2-yl) pyridine

A round bottom flask containing 3- (2,5-dihydrofuran-2-yl) -2-fluoropyridine (170 mg, 1 mmol) and 10% Pd / C in methanol (3.1 mL) is evacuated under vacuum and filled with hydrogen with the aid of a flask. The reaction is stirred overnight at ambient temperature and filtered over celite. The solvent is evaporated in vacuo to give 148 mg of 2-fluoro-3- (tetrahydrofuran-2-yl) pyridine. MS (m / z): 168 (M + 1). 3-Methyl-1- (3- (tetrahydrofuran-2-yl) pyridin-2-yl) -1H-pyrazole-4-carbaldehyde To a test tube with screw cap containing 2-fluoro-3- tetrahydrofuran-2-yl) pyridine (148 mg, 0.88 mmol) in dimethylformamide (2.2 mL) is added 3-methyl-1H-pyrazole-4-carbaldehyde (81 mg, 0.74 mmol) and carbonate of potassium (152 mg, 1.1 mmol). The reaction tube is quickly sealed (note: possible pressure buildup, use a safety shield) and stirred in a pre-agitated oil bath at 110 ° C for 18 h with the aid of a magnetic stirrer. The mixture is diluted with water and extracted with ethyl acetate. The organic layer is separated, dried over magnesium sulfate, filtered and the solvent evaporated in vacuo. The resulting residue is purified on silica gel using Isco normal phase chromatography eluting with hexane: acetone (5 to 20% gradient in acetone) to give 116 mg of the title compound. MS (m / z): 258 (M + 1). 60 ΡΕ2501703

Preparation 45: 3- (2-Bromo-pyrid-3-yl) isoxazole

1. 2-Bromopyridine-3-carbaldehyde oxime To a solution of 2-bromopyridine-3-carbaldehyde (2.04 g) in ethanol (25 mL) is added hydroxylamine hydrochloride (948 mg, 13.16 mmol) and ethanoate (1.09 g, 13.16 mmol). The mixture is stirred at room temperature overnight. The solvent is evaporated under reduced pressure and the residue is partitioned between dichloromethane and water. The organic layer is separated, dried over sodium sulfate, filtered and the solvent evaporated in vacuo to give 2 g of 2-bromopyridine-3-carbaldehyde oxime. MS (m / z): 201/203 (M + 1 / M + 3) 2. To a stirred suspension of 2-bromopyridine-3-carbaldehyde oxime (400 mg, 1.99 mmol) in dichloromethane (3.98 mL) at 0 ° C, pyridine (201.13 μ, 2.49 mmol) was added portionwise followed by N-chlorosuccinimide (337.19 mg, 2.49 mmol). Then, trimethylsilyl) acetylene (350.60 μ, 2.49 mmol) is added followed by triethyl-61α2501703 amine (346.68 æL, 2.49 mmol). The mixture is stirred at 0 ° C to room temperature. After two hours, more pyridine (321.81 æL, 3.98 mmol) and IV-chlorosuccinimide (539.51 mg, 3.98 mmol) are added and the mixture is stirred at room temperature for 16 hours. The reaction mixture is then quenched with saturated ammonium chloride solution. The layers are separated and the organics are dried over magnesium sulfate, filtered and concentrated under reduced pressure to afford [3- (2-bromo-3-pyridyl) isoxazol-5-yl] trimethylsilane in quantitative yield. To a solution of [3- (2-bromo-3-pyridyl) isoxazole (0.8 g) in dichloromethane -5-yl] -trimethylsilane (620 mg, 2.09 mmol) in methanol (18 mL) is added potassium carbonate (29.12 mg, 208.59 pmol) in one portion. The mixture is stirred at room temperature for 24 h. The solvent is removed in vacuo. The crude product is partitioned between ethyl acetate and saturated ammonium chloride solution. The organics are washed with water and dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product is purified by normal Isco phase chromatography eluting with hexane / ethyl acetate from 66/33 to 50/50 to give 140 mg of the title compound as a white solid. MS (m / z): 225/227 (M + 1 / M + 3) 62 ΡΕ2501703

Preparation 46: 1- (3-Chloro-2-pyridyl) -3-methyl-pyrazole-4-carbaldehyde

H

To a solution of 3-methylpyrazole-4-carbaldehyde (0.5 g, 4.5 mmol) in dry dimethylformamide (15 mL) is added potassium carbonate (2.5 g, 18 mmol) at room temperature . The reaction mixture is stirred at 40 ° C for 30 min and then 1-fluoro-2-chloropyridine (0.77 g, 6 mmol) is added. The reaction mixture is heated at 70 ° C for 16 h. Upon completion, the reaction mixture is cooled to room temperature, diluted with water and then extracted with ethyl acetate. The organic layer is dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude mixture is purified by silica gel chromatography eluting with hexane / ethyl acetate (75:25, 65:35) to give 0.525 g (52%) of the titled compound. MS (m / z): 222 (M + 1).

Preparation 47: 1- (3-Cyclopropyl-2-pyridyl) -3-methyl-pyrazole-4-carbaldehyde 63 ΡΕ2501703 1. 1- (3-Bromo-pyridin-2-yl) -3- To a solution of 3-methyl-pyrazole-4-carboxylic acid ethyl ester (2.0 g, 13 mmol) in dry dimethylformamide (30 mL) is added potassium carbonate (7.9 g, 2 g, 52 mmol) at room temperature. The reaction mixture is stirred at 40 ° C for 30 min and then 1-fluoro-2-bromopyridine (3.0 g, 17 mmol) is added. The reaction mixture is stirred at 70 ° C for 4 h. The reaction mixture is cooled to room temperature, diluted with water and then extracted with ethyl acetate. The organic layer is dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude mixture was purified by silica gel chromatography eluting with hexane / ethyl acetate (85:15, 80:20) to give 3.2 g (79%) of 1- (3-bromo-3- pyridin-2-yl) -3-methyl-pyrazole-4-carboxylic acid. MS (m / z): 312 (M + 3), 310 (M + 1). 2. 1- (3-Cyclopropyl-pyridin-2-yl) -3-methyl-pyrazole-4-carboxylic acid ethyl ester

A mixture of 1- (3-bromo-pyridin-2-yl) -3-methyl-pyrazole-4-carboxylic acid ethyl ester (1.5 g, 5.5 mmol), potassium phosphate gt; 19.2 mmol), cyclopropyl boronic acid (0.7 g, 8.2 mmol) and toluene / water (2: 1, 15 mL) is degassed with nitrogen and tricyclohexylphosphene (0.153 g, , 54 mmol) and palladium acetate (0.061 g, 0.27 mmol). The mixture is degassed 64 ΡΕ2501703 again with nitrogen and the reaction vessel is sealed and heated at 100 ° C for 16 h. The reaction mixture is concentrated, diluted with water and extracted with ethyl acetate. The organic layer is dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude mixture is purified by silica gel chromatography eluting with hexane / ethyl acetate (85:15) to afford 1.6 g (93%) of 1- (3-cyclopropyl-pyridin- 2-yl) -3-methyl-pyrazole-4-carboxylic acid. MS (m / z): 272 (M + 1). To a stirred suspension of lithium aluminum hydride (0.44 g, 11.7 mmol) in dichloromethane (3 mL) in dry tetrahydrofuran (25 mL) is added a solution of 1- (3-bromo-pyridin-2-yl) -3-methylpyrazole-4-carboxylic acid ethyl ester (1.6 g, 5.8 mmol ) in dry tetrahydrofuran (5 mL) under a nitrogen atmosphere at -78 ° C. After complete addition the reaction mixture is warmed to room temperature and then stirred overnight. The reaction mixture is quenched with 1N sodium hydroxide then filtered through celite. The organic layer is separated and the aqueous layer is extracted with ethyl acetate. The combined organic layer is washed with water and brine, dried over sodium sulfate, filtered and concentrated to afford 1.1 g (84%) of [1- (3-cyclopropyl-pyridin-2-yl) -3-methyl- pyrazol-4-yl] -methanol. MS (m / z): 230 (M + 1). 65 ΡΕ2501703 4. To a solution of [1- (3-cyclopropyl-pyridin-2-yl) -3-methyl-pyrazol-4-carbaldehyde 4-yl] -methanol (1.1 g, 5.0 mmol) in dry dichloromethane (15 mL) is added pyridinium chlorochromate (1.2 g, 5.5 mmol) under a nitrogen atmosphere. The reaction mixture is stirred at room temperature for 2 h and then filtered through celite. The filtrate is concentrated under reduced pressure. The crude mixture is purified on silica gel eluting with hexane / ethyl acetate (90:10, 80:20) to afford 0.2 g (20%) of the titled compound. MS (m / z): 228 (M + 1).

Preparation 48: 4 - [(2-Bromo-3-pyridyl) methyl] morpholin-3-one

Br

To a solution of 2-bromo-3-bromomethyl-pyridine (0.68 g, 2.36 mmol) in dimethylformamide (10 mL) are added sodium hydride (0.11 g, 60% suspension, 2.82 mmol ), morpholin-3-one (0.20 g, 1.97 mmol) and tetrabutylammonium iodide (catalytic) at 0 ° C. The mixture is stirred at room temperature for 16 hours. After completion, the reaction mixture is partitioned between ethyl acetate (25 mL) and water (25 mL). The aqueous layer is extracted with ethyl acetate (2 x 25 mL) and the combined organic extracts are dried over sodium sulfate and concentrated in vacuo. The crude material is purified by column chromatography on silica gel eluting with hexane / ethyl acetate (55:45) to give 0.35 g (66%) of the title compound. MS (m / z): 271/273 (M + 1, M + 3).

The compounds of Preparations 49-50 are prepared essentially as described in Preparation 48 from the corresponding lactam reagent.

Prep. No. 1 - [(2-Bromo-3-pyridyl) methyl] -pyrrolidin-2-one Br ocher 255 255/257 (M + 1, M + H +) + 3) 50 1 - [(2-Brhenio-3-pyridyl) methyl] piperidin-2-one Br 0 συ 59 269/271 (M + 1 / M + 3)

Preparation 51: Ethyl 1- (3-chloro-2-pyridyl) -4-formyl-pyrazole-3-carboxylate

EtO

To a solution of ethyl 4-formyl-pyrazole-3-carboxylate (1.70 g, 10.1 mmol) and 3-chloro-2-fluoropyridine (1.99 g, 15.1 mmol) in dimethylformamide (20 mL ) is added potassium carbonate (2.80 g, 20.2 mmol) and the mixture is stirred at 100 ° C for 16 hours. After completion, the reaction mixture is partitioned between water (25 mL) and ethyl acetate (25 mL). The aqueous layer is extracted with ethyl acetate (3 x 25 mL) and the combined organic extracts are dried over sodium sulfate, concentrated in vacuo and purified by silica gel column chromatography eluting with hexane / ethyl acetate (50 mL). : 50) to afford 1.5 g (54%) of the title compound. MS (m / z): 280 (M + 1).

Preparation 52: Ethyl 1- (3-methyl-2-pyridyl) -4-formyl-pyrazole-3-carboxylate

A mixture of ethyl 1- (3-chloro-2-pyridyl) -4-formyl-pyrazole-3-carboxylate (1.0 g, 3.5 mmol), trimethyl boroxine (0.44 g, , 50 mmol) and potassium carbonate (1.4 g, 10.1 mmol) in dioxane (18 mL) and water (2 mL) is degassed with nitrogen gas for 10 min and then palladium-triphenylphosphine , 40 g, 0.3 mmol) and stirred at 110 ° C for 16 h. Upon completion, the reaction mixture is partitioned between dichloromethane (50 mL) and water (25 mL). The aqueous layer is extracted with dichloromethane (2 x 25 mL) and the combined organic extracts are dried over sodium sulfate and concentrated in vacuo. The crude mixture is purified by column chromatography on silica gel eluting with hexane / ethyl acetate (70:30) to give the title compound (0.12 g, 13%). MS (m / z): 260 (M + 1). 68 ΡΕ2501703

[2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1- (L) -L yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol

To a solution of 2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) -3-methylpyrazol-1-yl] pyridine-3-carbaldehyde (12.50 g, 26.10 mmol) in dichloromethane (125 mL) at 0 ° C is added sodium borohydride (0.395 g, 44 mmol) and methanol (37.50 mL). The ice bath is removed and the reaction is stirred at room temperature for 30 min. Water (50 mL) is added and the mixture is concentrated to one-half volume resulting in a sticky white solid precipitate. Dichloromethane (100 mL) is added and the biphasic mixture is separated. The organic layer is washed with water (50 mL), dried over sodium sulfate and concentrated to one-half volume (about 100 mL). Methyl tert-butyl ether (100 mL) is added and the solution is concentrated to provide a precipitate. The solid is filtered and dried under vacuum to give 12 g of [2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'- piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol. MS (m / z): 481 (M + 1). The free base of the title compound (0.48 g, 1 mmol) is dissolved in 3 mL of methanol and a solution of (L) -tartaric acid (0.15 g, 1 mmol) in methanol is added. The mixture is stirred for ten minutes. The solvent is evaporated and the residue is dried in vacuo overnight to afford [2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol in quantitative yield. MS (m / z): 481 (M + 1).

Example 2: 2-Chloro-4,4-difluoro-1 '- [[1- [3- (4H-imidazol-2-yl) -2-pyridyl] -3-methyl-pyrazol-4-yl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine]

A solution of 40% glyoxal in water (4.69 mL, 40.71 mmol) and 2N ammonia in methanol (54.29 mL, 108.57 mmol) is added to a suspension of 2- [4 - [(2 chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -pyridine -3-carbaldehyde (6.50 g, 13.57 mmol) in methanol (10 mL). The reaction mixture is stirred for five minutes at 0øC and then at 40øC for 1 h. Dichloromethane (10 mL) is added to a slurry dissolving almost all of the material. The mixture is stirred for 4 h resulting in a light brown solution. The 70 ΡΕ2501703 solution is stirred for 8 h at 40øC. 40% glyoxal in water (2.34 mL, 20.36 mmol) and 2N ammonia in methanol (27.14 mL, 54.29 mmol) are added and the mixture is stirred at 40 ° C for 15 h. The reaction mixture is concentrated to approximately 100 mL and poured into ice / water (400 mL) and a beige solid precipitate forms. The solid is filtered, washed with water and dried under vacuum (7 g). The solid is combined with other raw materials prepared essentially by the same method (10 g). The combined solid is dissolved in dichloromethane and filtered through a pad of silica gel eluting with dichloromethane / methanol (96/4). Fractions containing product are combined and concentrated to give a light yellow solid (11 g). The solid is extracted with 10% hexane / dichloromethane under reflux and the filtrate is concentrated. The solid is further purified via silica gel chromatography eluting with 2N ammonium hydroxide in methanol / dichloromethane mixtures to obtain 6.5 g of the title compound. MS (m / z): 518 (M + 1).

Example 3: 2-Fluoro-1 '- [[1- [3- (1H-imidazol-2-yl) -2-pyridyl] -3-methylpyrazol-4-yl] methyl] spiro [ 5-dihydrothieno [2,3-c] pyran-7,4'-piperidine]

This compound is prepared essentially as 71 ΡΕ 2505070 described in Example 2 using 2- [4 - [(2-fluoro-spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] yl) methyl] -3-methyl-pyrazol-1-yl] pyridine-3-carbaldehyde. The residue is purified by standard Isco phase chromatography to give 32% of the title compound: mass spectrum (m / z): 465 (M + 1).

Example 4: 5- (2- (4 - ((2'-Chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3- pyran-1-yl) methyl) -3-methyl-1H-pyrazol-1-yl) pyridin-3-yl) oxazole

To a screw cap test tube containing 2- (4 - ((2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [ 3-c] pyran] -1-yl) methyl) -3-methyl-1H-pyrazol-1-yl) nicotinaldehyde (0.120 g, 0.25 mmol) in methanol (2.5 mL) is added potassium carbonate 0.104 g, 0.75 mmol) and p-toluenesulfonylmethyl isocyanide (0.054 g, 0.28 mmol) under nitrogen. The reaction tube is quickly sealed (note: possible pressure buildup, use a safety shield) and is shaken in an oil bath preheated at 65 ° C for 3 h with the aid of a magnetic stirrer. The mixture is diluted with methanol and purified using a 2 g SCX cartridge. The solvent is evaporated in vacuo. The resulting residue is purified by flash chromatography on 72/2501703 Isco normal phase eluting with hexane / ethanol (5-30%) to give 0.093 g of the title compound. MS (m / z): 518 (M + 1)

Example 5: 2-Chloro-4,4-difluoro-1 - [[1- (3-fluoro-2-pyridyl) -3-methyl-pyrazol-4-yl] methyl] spiro [5H -thieno [2,3-c] pyran-7,4'-piperidine]

To a solution of 2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] (19.99 g, 64.33 mmol) in 1,2 dichloroethane (120 mL) is added 1- (3-fluoro-2-pyridyl) -3-methyl-pyrazole-4-carbaldehyde (12 g, 58.48 mmol) and the mixture is stirred at room temperature for 15 min. Then powdered sodium triacetoxyborohydride (18.59 g, 87.72 mmol) is added (internal temperature 25-35ø) and the resulting suspension is stirred at room temperature for 15 h. A mixture of saturated aqueous sodium bicarbonate solution on ice (200 mL) is added portionwise with stirring. The phases are separated. The aqueous phase is extracted with methyl tert-butyl ether (30 mL). The combined organic layers are washed with water (50 mL) and 50% brine (50 mL), dried over sodium sulfate and concentrated to give a thick oil which is purified via silica gel chromatography using dichloromethane / methanol mixtures as the eluent to give 18 g of a thick 73 ΡΕ2501703 oil which is triturated with hexane and 10% methyl tert-butyl ether / hexane to obtain 16.5 g of 2-chloro-4,4-difluoro-1 ' - [[1- (3-fluoro-2-pyridyl) -3-methyl-pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine]. MS (m / z): 469 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 469 (M + 1).

The compounds of Examples 6-15 are prepared essentially as described in Example 5 using the corresponding aldehyde.

(I /): 1- [2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [ 2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] -N, N -dimethyl-methanamine FF (M + 1) 23 523 (M + 1) 7 (t) -N - [[2- [4-Amino- - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1 -yl] -3-pyridyl] methyl] acetamide 2-fluoro-1 '- [[1- [3 (R) -cyclohexanecarboxylate - (methoxymethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methyl] spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] (2-fluoro-2-pyridyl) -3-methyl-pyrazol-2-yl) 4-yl] methyl] spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] (continuation)

Ex. Chemical name Structure Yield% Physical data: MS (m / z) (L) -Total 2,4,4-trifluoro-1 '- [[1- [3-FF S " > (methoxymethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-y and 72,479 (MEL) piperidine] - > (3-fluoro-2-pyridyl) -3- (methoxymethyl) pyrazol-3-yl] -piperidine-4-carboxylic acid tert- 4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] 64 499 (M + 1) (L) -2,4,4-trifluoro- - [[1- (3-t-Fluoro-2-pyridyl) -3-methyl-pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7.4 2-Chloro-4,4-difluoro-1 '- [[3-O-y-1' Methyl-1- (3-morpholino-2-pyridyl) pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] 60 536 ( 2-chloro-1 '- [[1- [3- (methoxymethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methyl] spiro [ 5-dihydrothieno [2,3-c] pyran-7,4'-piperidine]) and 2-chloro-1 '- [[ 1- [3- (cyclopropoxymethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methyl] -1H- 4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] The reactions are carried out using tetrahydrofuran as solvent. 75 ΡΕ2501703

Example 16: 2-Chloro-4,4-difluoro-1 '- [[3-methyl-1- (3-tetrahydrofuran-2-yl-2-pyridyl) pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine]

To a screw-capped test tube containing a mixture of 3-methyl-1- (3- (tetrahydrofuran-2-yl) pyridin-2-yl) -1H-pyrazole-4-carbaldehyde (192 mg, 75 mmol), 2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran (209 mg, 0.75 mmol) and acetic acid (43 æL, 0.75 mmol) in 1,2-dichloroethane (2.6 mL) is added sodium triacetoxyborohydride (238 mg, 1.1 mmol). The reaction tube is sealed and stirred at room temperature for 18 h with the aid of a magnetic stirrer. The mixture is diluted with methanol and purified using a 10 g SCX cartridge and the solvent evaporated in vacuo. The resulting residue is further purified by standard Isco phase chromatography eluting with hexane: acetone (5 to 30% gradient in acetone) to give 113 mg of the title compound as a racemic mixture. MS (m / z): 521 (M + 1). The enantiomeric resolution is carried out under SCR (supercritical fluid chromatography) conditions used as Chiralpak AD-H® (Chiral Technologies, Inc., West Chester, Pennsylvania, USA) as the stationary phase and carbon dioxide / methanol / dimethylethylamine (0.2%) as the mobile phase. The first eluent is the desired enantiomer.

Example 17: 2-Chloro-4,4-difluoro-1 '- [[1- (3-fluoro-2-pyridyl) -3- (isopropoxymethyl) pyrazol-4-yl] methyl tartrate (L) ] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine]

To a solution of 2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] (170 mg, 0.61 mmol) in dichloromethane mL), 1- (3-fluoro-2-pyridyl) -3- (isopropoxymethyl) pyrazole-4-carbaldehyde (192 mg) is added. The mixture is stirred 10 min at room temperature. Then, sodium triacetoxyborohydride (268.3 mg) is added, and the reaction is stirred at room temperature overnight. The mixture is diluted with dichloromethane and quenched slowly with sodium bicarbonate (saturated solution). The organic phase is then extracted with more dichloromethane, decanted, dried over magnesium sulfate and the solvent evaporated under reduced pressure. The residue is purified by reverse phase HPLC to give 67.9 mg of 2-chloro-4,4-difluoro-1 '- [[1- (3-fluoro-2-pyridyl) -3- (isopropo -oxymethyl) pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine]. MS (m / z): 527 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 527 (M + 1).

Example 18: 2-Chloro-4,4-difluoro-1 '- [[1- [3- (methoxymethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine]

To a test tube with screw cap is added copper (I) iodide (1.15 g, 6.02 mmol), 2-chloro-4,4-difluoro-1 '- [(3-methyl- pyrazol-4-yl) methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] (15 g, 40.12 mmol), potassium carbonate (11.76 g, 84 mmol) , 26 mmol), 15 mL of toluene (previously bubbled with nitrogen for 20 minutes) and a stir bar. The reaction mixture is bubbled with nitrogen for a further 10 min and then 2-bromo-3-methoxymethyl-pyridine (10.54 g, 52.16 mmol), and trans-N, N'-dimethylcyclohexane-1 , 2-diamine (1.90 mL, 12.04 mmol). The reaction tube is quickly sealed (caution: possible pressure increase, use a safety shield), stirred 78 ΡΕ2501703 at room temperature for 5 min and immersed in an oil bath preheated at 115 ° C for 24 h. The sample is cooled to room temperature, diluted with ethyl acetate and filtered through celite. The solvent is evaporated in vacuo. The residue is purified by standard Isco phase chromatography using hexane / ethyl acetate (30-70%). The desired fractions are collected and evaporated. Some impure fractions (<2g) are repurified by Isco normal phase chromatography using hexane / ethyl acetate (20-60% in ethyl acetate) as the eluent. All fractions containing final compound are combined and evaporated to dryness in vacuo. The residue is dissolved in dichloromethane and washed with 10% aqueous ammonium hydroxide solution to remove copper residues. The organic solvent is evaporated and the compound is dried overnight to give 13 g of 2-chloro-4,4-difluoro-1 '- [[1- [3- (methoxymethyl) -2-pyridyl] - 3-methyl-pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4 '-piperidine]. MS (m / z): 495 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 495 (M + 1).

The compounds of Examples 19-35 are prepared essentially as described in Example 18 from the corresponding 2-bromo-3-substituted pyridine (examples 19-32 & 34-35) or 2-iodo-3-substituted pyridine ( example 33). 79 ΡΕ2501703 Εχ. No. 2-Chloro-1 '- [[1- [3- (ethoxymethyl) -2-pyridyl] -3-methylpyrazole-2-carboxylic acid tert- methyl-pyrazol-4-yl] methyl] -4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] 2-chloro-1 '- [[1- [3- (cyclopropylmethoxymethyl) -2-pyridyl] -3-methyl-pyrazol-4-yl] methyl] -4- 4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine]; 2-Chloro-4,4-difluoro-1 '- [[1- [3- (isopropoxymethyl) -2-pyridyl] -3-methyl-pyrazole-4-carboxylate -yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] (M + 1) 22 (L) - 2-Chloro-4,4-difluoro-1 '- [[3-methyl-1- (3-methyl-2-pyridyl) pyrazol-4-yl] methyl] spiro [5H- c] pyran-7,4'-piperidine] -L-Amino-3-methyl-1 - [(3-chlorophenyl) - [3- (pyrazol-1-ylmethyl) -2-pyridyl] pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] 2-chloro-4,4-difluoro-1 '- [[3-methyl-1- [3- (triazol-2-ylmethyl) -2- pyridyl] pyrazol-4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] (Continued) )

Ex. No. Chemical name Structure Yield (%) Physical data: MS (m / z) (L) -2-Chloro-4,4-difluoro-1 '- [[3-methyl-1- [3- (1,2,4-triazol-1-ylmethyl) -2-pyridyl] pyrazol-4-one [N, N -dimethylamino] methyl] spiro [5H-thieno [2,3- C] pyran-7,4'-piperidine] 2-fluoro-1 '- [[3-Cyclopropyl] -methyl-1- [3- (1 (MEL) yl] methyl] spiro [4,5-dihydrothieno [2,3-c] pyridin-4-yl] pyrimidine-4-carboxylic acid tert-butyl ester (2-chloro-4,4-difluoro-1 '- [[1- [3- (imidazole (MEL) 4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4apos; -piperidine] pyridin-3-yl] & 4-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-o-28 (c) pyran-7,4- (1-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] acetonitrile 2 - [[2 (R) - [4 - [(2-chloro- [4,4-difluoro-spiro [5H-thieno [2,3-b] pyridin-7,4apos; -1 ', 13 524 (MEL) yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] oxy] acetamide (L) rO '0. N-methyl-1- [3 - [(2-methylimidazol-1-yl) methyl] -2-pyridyl] pyrazol-4- 0 57 493 (MEL) yl] methyl] spiro [4,5-dihydroxy-N, N -dimethyl- [2,3- c] pyran-7,4'-piperidine] N, H 81 ΡΕ2501703 (cont.)

Example 36: 2-Chloro-4,4-difluoro-1 '- [[1- (3-methoxy-2-pyridyl) -3-methylpyrazol-4-yl] methyl] spiro [5H-thieno [ 2,3-c] pyran-7,4'-piperidine] 82 ΡΕ2501703

To a test tube with screw cap is added copper (I) iodide (23.74 mg, 0.125 mmol), 2-chloro-4,4-difluoro-1 '- [(3-methyl-1H-pyrazol-4 yl) methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] (233.00 mg, 0.623 mmol), potassium carbonate (182.71 mg, 1.31 mmol) , toluene (2 mL) (previously bubbled with nitrogen for 20 min) and a stir bar. The reaction mixture is bubbled with nitrogen for 20 minutes and then 2-iodo-3-methoxypyridine (302.01 mg, 1.25 mmol) and trans-N, N'-dimethylcyclohexane-1,2-diamine (39.31 æl, 0.25 mmol). The reaction tube is quickly sealed (note: possible pressure increase, use a safety shield) and immersed in an oil bath preheated at 110 ° C for 24 hours with the aid of a magnetic stirrer. The mixture is then poured into SCX column (25 g) and eluted with methanol and then 2N ammonia solution in methanol. The basic fraction is concentrated and the resulting residue is purified by basic HPLC to give 162 mg of the title compound. MS (m / z): 481 (M + 1).

Example 37: 2-Chloro-4,4-difluoro-1 '- [[1- (3-isoxazol-3-yl-2-pyridyl) -3-methyl-pyrazol-4-yl] methyl] spiro [5H- thieno [2,3-c] pyran-7,4'-piperidine] 83 ΡΕ2501703

This compound is essentially prepared as described in Example 36 using 2-chloro-4,4-difluoro-1 '- [(3-methyl-1H-pyrazol-4-yl) methyl] spiro [5H-thieno [2,3- -c] pyran-7,4'-piperidine] and 3- (2-bromo-pyrid-3-yl) isoxazole as the starting material. The residue is purified by standard Isco-phase chromatography (ethyl acetate is used as eluent) to afford 45% of the title compound: mass spectrum (m / z): 518 (M + 1)

Example 38: 2- (4 - ((2'-Chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3- yl) methyl) -3-methyl-1H-pyrazol-1-yl) pyridin-3-yl) propan-2-ol

To a screw cap test tube containing 2- (2-bromo-3-pyridyl) propan-2-ol (520 mg, 2.41 mmol) in dry dimethylformamide (3.2 mL) are added under nitrogen 4 - ((2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine- 84 ΡΕ2501703 4,7 '-thieno [2,3-c] pyran] -1-yl ) (R) - (-) - N, N'-methyl-3-methyl-1H-pyrazole (600 mg, 1.6 mmol) dimethyl-1,2-cyclohexanediamine (68 mg, 480 mmol) and cesium carbonate (1040 mg, 3.2 mmol). The reaction tube is quickly sealed (note: possible pressure increase, use a safety shield) and stirred in an oil bath preheated at 110 ° C for 16 h with the aid of a magnetic stirrer. The mixture is diluted with water and extracted with ethyl acetate. The organic layer is separated, dried over magnesium sulfate, filtered and the solvent evaporated in vacuo. The resulting residue is purified by silica gel using standard Isco phase chromatography eluting with hexane: ethanol (2 to 15% gradient in ethanol) to give 310 mg of the title compound. MS (m / z): 509 (M + 1)

Example 39: 1- [2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'- piperidinyl] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] -N-methyl-methanamine

The free base of the title compound is prepared essentially as described in Preparation 7 using 2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-85β] -250,170,37 , 4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] pyridine-3-carbaldehyde and methylamine in 51% yield. MS (m / z): 494 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 551 (M + 1).

The compounds of Examples 40-44 are prepared essentially as described in Example 39 from the corresponding aldehyde and amine. (Example 43 is prepared using 1,2-dichloroethane as solvent)

Example 4: N - [[2- [4 - [(2-chloro-4,4-difluoro-spiro [1] 5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methyl] propan-2-amine (M +) 34 522 (MEL) 41 (L) -T- [[2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3- c] pyran -7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methylamino] acetonitrile, 2 - [[2- [4 - [(2-Fluorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl ) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methylamino] acetonitrile. r N-y H C-1 27 453 (M + 1) 86 ΡΕ2501703

Example 45: 2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1 ' -yl) methyl] -3-methyl-pyrazol-1-yl] pyridine-3-carbaldehyde oxime

To a solution of 2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] 3-carbaldehyde (0.210 g, 0.438 mmol) in ethanol (3 mL) are added hydroxylamine hydrochloride (0.038 g, 0.526 mmol) and sodium ethanoate 87 ΡΕ2501703 (0.044 g , 0.526 mmol) and the mixture is stirred under reflux for 1 h. The solvent is evaporated and the residue is extracted with ethyl acetate and water. The organic layer is separated, dried over magnesium sulfate, filtered and the solvent is evaporated in vacuo. The residue is purified by reverse phase basic HPLC to give 0.110 g of 2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4 '-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] pyridine-3-carbaldehyde oxime. MS (m / z): 494 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 494 (M + 1).

The compounds of Examples 46-47 are prepared essentially as described in Example 45 from the corresponding aldehyde and hydroxylamine.

88 ΡΕ2501703

Example 48: [2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1- (L) -tartrate) '-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanamine

1 - [[2- [4- [1- (2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl ) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methyl] isoindoline-1,3-dione

Diisopropyl azodicarboxylate (0.105 mL, 0.54 mmol) is added to a solution of [2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-1-yl] -3-pyridyl] methanol (0.173 g, 0.36 mmol), phthalimide (0.079 mg, 0.7 mmol) , 54 mmol) and triphenylphosphine (0.142 g, 0.54 mmol) in toluene (3 mL) at 0 ° C. The mixture is stirred at room temperature overnight. The solvent is removed and the residue is first purified using a 2 g SCX cartridge after evaporation of the 2N ammonia fraction in methanol by normal Isco phase chromatography eluting with hexane / ethanol (3% - 30%) to give 0.201 g of 2 - [[2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) -3-methyl-pyrazolo [3,4-b] pyrrol-1-yl] -3-pyridyl] methyl] isoindoline-1,3-dione. MS (m / z): 610 (M + 1). [2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1 ' -yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanamine To a flask containing 2 - [[2- [4 - [(2-chloro-4,4-difluoro-spiro [5H 2-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methyl] isoindoline-1,3-thieno [2,3- c] pyran-7,4'- dione (0.201 g, 0.33 mmol) and 24 μl water in ethanol (2.5 mL) is added hydrazine monohydrate (0.04 g, 0.79 mmol). The reaction mixture is refluxed for 2.5 h and then diluted with methanol and purified using a 2 g SCX cartridge. After evaporation of the 2N ammonia in methanol fraction, the resulting product is purified by Isco normal phase chromatography eluting with ethanol and a 15% ammonium hydroxide solution (7N in methanol) in ethanol (25-90% gradient of eluent) yielding 129 mg of the title compound. MS (m / z): 480 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 480 (M + 1).

Example 49: 2- [4 - [(2-Fluorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl ] -3-methyl-pyrazol-1-yl] pyridine-3-carboxamide 90 ΡΕ2501703

1- [4 - [(2-Fluorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazole -1-yl] pyridine-3-carbonitrile:

This compound is essentially prepared as described in Preparation 7 using 2-fluoro-spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] and 2- (4-formyl-3-methyl pyrazol-1-yl) pyridine-3-carbonitrile. The residue is purified through SCX to give 2- [4 - [(2-fluoro-spiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl ] -3-methyl-pyrazol-1-yl] pyridine-3-carbonitrile in 32% yield. MS (m / z): 424 (M + 1). 2. (L) -2- [4 - [(2-Fluorospiro [4,5-dihydro-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] pyridine-3-carboxamide To a solution of 2- [4 - [(2-fluoro-spiro [4,5-dihydrothieno [2,3- c] 7-yl] pyridine-3-carbonitrile (0.71 mmol, 0.30 g) in dimethylsulfoxide (7.5 mL) , potassium carbonate (0.049 g, 0.35 mmol) is added and the mixture is cooled to 0-5øC before adding peroxide solution of 91 ΡΕ2501703 hydrogen in 33% water (0.39 mL, 3.78 mmol). The mixture is stirred at room temperature for 2 h. Then water is carefully added and the reaction mixture is extracted with ethyl acetate. The organic layer is decanted, dried over magnesium sulfate and the solvent is evaporated under reduced pressure. The crude product is purified by Isco normal phase chromatography using dichloromethane / methanol mixtures as eluent to give 0.252 g which are further purified by reverse phase HPLC to give 0.198 g of 2- [4 - [(2-fluoro-spiro [ 4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] pyridine-3-carboxamide. MS (m / z): 442 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 442 (M + 1).

Example 50: 2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1 ' -yl) methyl] -3-methyl-pyrazol-1-yl] pyridine-3-carboxamide

The title compound is prepared essentially as described in Example 49 using 2-chloro-4,4-difluoro-9β, 252β-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] and 2 - (4-formyl-3-methyl-pyrazol-1-yl) pyridine-3-carbonitrile as starting materials in 15% overall yield (22% yield in the first step and 66% yield in the second step). MS (m / z): 494 (M + 1).

Example 51: 2- [2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'- piperidinyl] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] acetamide

To a solution of 2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl ) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] acetonitrile (170 mg, 346.96 mmol) in dimethylsulfoxide (4 mL) is added potassium carbonate (23.98 mg, 173.48 mmol) ) and the mixture is cooled to 0-5 ° C. 33% Hydrogen peroxide solution in water (189.54 æL, 1.84 mmol) is added. The mixture is stirred at room temperature for 16 h and then further 33% hydrogen peroxide solution in water (189.54 æl, 1.84 mmol) and potassium carbonate (23.98 mg, 173.48 pmol) are added. Water is added and the mixture is extracted with ethyl acetate. The organic layer is separated, washed with brine, dried over magnesium sulfate and the solvent evaporated in vacuo. The compound is purified by standard Isco phase chromatography using dichloromethane and 2N ammonia in methanol as eluents (1.5 to 6% in 2N ammonia in methanol) to afford 19% of the free base of the title compound. MS (m / z): 508 (M + 1). The tartrate salt is prepared essentially as described in Example 1. MS (m / z): 508 (M + 1).

Example 52: [2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'- piperidin] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methyl

Methyl isocyanate (5.80 æl, 0.096 mmol) is added to a solution of 2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7 , 4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol (0.042 g, 0.087 mmol) in dichloromethane (1 mL) at room temperature. After 94 ΡΕ2501703 h and no evolution of the reaction through LC / MS, triethylamine (12.17 æl, 87.32 pmol) and methyl isocyanate (5.8 æl, 0.096 mmol) are added and the reaction is stirred one day to the next. Further triethylamine (12.17 μL, 87.32 μmol) and methyl isocyanate (5.8 μL, 0.096 mmol) are added and after 24 h the solvent is evaporated and the residue is purified by standard Isco phase chromatography with dichloromethane / 2N ammonium hydroxide in methanol (0 to 5% in 2N ammonium hydroxide in methanol) as eluent to afford 0.041 g of [2- [4 - [(2-chloro-4,4-difluoro- spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methyl ester. MS (m / z): 538 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 538 (M + 1).

Example 53: [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl ) methyl] -1- (3-fluoro-2-pyridyl) pyrazol-3-yl] methanol

95 ΡΕ2501703 1. 4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1- (3-fluoro-2-pyridyl) pyrazole-3-carboxylate To a solution of ethyl 1- (3-fluoro-2-pyridyl) -4-formyl-pyrazole-3-carboxylate (342, 5 mg) in 1,2-dichloroethane (6.7 mL), 2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4 '-piperidine] ( 280 mg, 1 mmol). The mixture is stirred for 10 min. Thereafter, sodium triacetoxyborohydride (442 mg) is added and the mixture is stirred overnight at room temperature. Sodium bicarbonate (saturated aqueous solution) is added and the organic phase extracted with dichloromethane, dried over magnesium sulfate and the solvent evaporated in vacuo. The crude product is purified by normal Isco phase chromatography eluting with ethyl acetate and hexane to give 450 mg of 4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c ] pyran-7,4'-piperidine] -1'-yl) methyl] -1- (3-fluoro-2-pyridyl) pyrazole-3-carboxylate. MS (m / z): 527 (M + 1). [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1- (3-fluoro-2-pyridyl) pyrazol-3-yl] methanol To a solution of 4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3- c ] pyran-7,4'-piperidine] -1'-yl) methyl] -1- (3-fluoro-2-pyridyl) pyrazole-3-carboxylate (250 mg, 0.47 mmol) in tetrahydrofuran (2.37 mL) under nitrogen atmosphere and cooled to 0 ° C, 1M lithium aluminum hydride in tetrahydrofuran (569.3 μL) is slowly added. The mixture is stirred at that temperature for 30 minutes. Then 22 μl of water are added and the cooling bath is removed and the mixture is stirred at room temperature for 5 min before adding 22 μl of 15% aqueous sodium hydroxide solution and 65 μl of water. The solids are filtered off and the solvent is evaporated. The crude product is purified by standard Isco phase chromatography using dichloromethane and methanol as eluent to give 141 mg of the title compound as the free base. MS (m / z): 485 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 485 (M + 1).

An alternative method to make the free base of Example 53: 1. 4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] - 1'-yl) methyl] -1H-pyrazole-3-carboxylate

To a suspension of 2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine (18.3 g, 65.42 mmol) in tetrahydrofuran (0.2 L) is added ethyl 4-formyl-1H-pyrazole-3-carboxylate (10 g, 59.47 mmol) and the mixture is stirred at room temperature for 10 min. Then, powdered sodium triacetoxyborohydride (16.39 g, 77.31 mmol) is added. The mixture is stirred at room temperature for 1.5 h. The reaction mixture is then poured into saturated aqueous sodium bicarbonate solution (200 mL). The phases are separated. The aqueous phase is basified with saturated NaHCO3 solution and extracted with ethyl acetate (100 mL). Combined organic layers are washed with saturated NaHCO3 solution, dried over sodium sulfate and concentrated to give 22.5 g of the title compound. MS (m / z): 432 (M + 1). 2. 4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1- (3- -fluoro-2-pyridyl) pyrazole-3-carboxylate

To a solution of 4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1H- pyrazole-3-carboxylate (22.5 g, 52.10 mmol) in dimethylformamide (225.00 mL) are added Potassium Carbonate (10.80 g, 78.14 mmol) and 2,3-difluoropyridine 98 ΡΕ2501703 (7.19 g, 62.52 mmol) and the resulting suspension is stirred at 60 ° C for 15 h. Then the reaction mixture is poured onto ice / brine (30 mL) and CH 2 Cl 2 (50 mL) is added to the resulting suspension. The solution is washed with H2O (2 x 50 mL). The organics are dried over sodium sulfate and concentrated to give a crude product which is purified via SiO2 filtration eluting with 2-propanol / CH2Cl2 mixtures to give 18 g of the title compound. MS (m / z): 527 (M + 1). 3. [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1- ( Fluoro-2-pyridyl) pyrazol-3-yl] methanol

To a solution of 4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] - Ethyl 1- (3-fluoro-2-pyridyl) pyrazole-3-carboxylate (2.6 g, 4.93 mmol) in tetrahydrofuran (26.00 mL) under N 2 atmosphere and cooled to -10 ° C , 1M Di-isobutyl aluminum hydride in toluene (23:68 mL, 23.68 mmol) is slowly added dropwise. Then the reaction mixture is stirred at this T for 15 min and then the cold bath is removed and the mixture is warmed to room temperature. Then, H2 O (100 mL) is added dropwise at -10 ° C. The resulting suspension is extracted with EtOAc (2 x 30 mL). The combined 99 ΡΕ2501703 are washed with H2 O (50 mL) and brine (50 mL), dried over Na2 SO4 and concentrated to give a pale brown oil which is dissolved in methyl t-butyl ether (10 mL) and crystallized white crystals. The suspension formed is stirred and hexane (10 mL) is added dropwise while stirring takes place. The solid is filtered to give 2 g of the title compound. MS (m / z): 485 (M + 1).

Example 54: 2- (4 - ((2'-Chloro-4 ', 4'-difluoro-4', 5'-dihydro-spiro [piperidine-4,7'-thieno [2,3- c] Pyran-1-yl) -3- (3-methyl-1,2,4-oxadiazol-5-yl) pyridine

1. 2- (4 - ((2'-Chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3- c] pyran] -1-yl) methyl) -3-methyl-pyrazol-1-yl) -nicotinic acid To a test tube with screw cap is added copper (I) oxide (11 mg, 0.08 mmol), 2-chloro-4,4-difluoro-1 '- [(3-methyl-1H-pyrazol-4-yl) methyl] spiro [5H-thieno [2,3- c] pyran-7,4'-piperidine] (300 mg, 0.8 mmol), cesium carbonate (523 mg, 1.6 mmol), dry dimethylformamide (1.6 mL) and a stir bar. The reaction mixture 100 ΡΕ2501703 is bubbled with nitrogen for 20 minutes and then methyl 2-iodopyridine-3-carboxylate (511 mg, 1.9 mmol) and trans-N, N'-dimethylcyclohexane-1,2 diamine (0.24 mmol, 34 mg). The reaction tube is quickly sealed (note: possible pressure increase, use a safety shield) and immersed in an oil bath preheated at 110 ° C for 16 hours with the aid of a magnetic stirrer. The mixture is then poured onto SCX column (10 g) and eluted with methanol and then 2N ammonia solution in methanol. The basic fraction is concentrated and the resulting residue is purified on silica gel using standard Isco phase chromatography eluting with hexane: ethanol (2 to 15% gradient in ethanol) to give 307 mg of 2- ( 4 - ((2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-yl) methyl ) -3-methyl-1H-pyrazol-1-yl) -nicotinic acid. MS (m / z): 309 (M + 1). 2. 2- (4 - ((21-Chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] - 1-yl) methyl) -3-methyl-1H-pyrazol-1-yl) -3- (3-methyl-1,2,4-oxadiazol-5-yl) pyridine To a solution of acetamide oxime , 2.1 mmol) in 7 mL of tetrahydrofuran, 139 mg of ground 4A molecular sieves and sodium hydride (2.1 mmol, 84 mg, 60% in mineral oil) are added under argon. The mixture is allowed to stir for 30 min at 50 ° C. After cooling to room temperature, 2- (4 - ((2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7- (309 mg, 0.6 mmol) and then the mixture is washed with dichloromethane and dried over sodium sulfate. heated at 50 ° C for 40 min. It is quenched after cooling with water and extracted with dichloromethane. After drying the organic phase with magnesium sulfate and evaporation of the solvent, the residue is diluted in methanol and purified using a 5 g SCX cartridge. The resulting residue is further purified by semipreparative MS reverse phase HPLC using an Xbridge column (5æm, 19x100 mm) and a gradient between 60 and 80% B in A in 5 min at a flow rate of 25 ml / min ( basic conditions of A: 20 mM ammonium bicarbonate pH 9 and B: acetonitrile) to give 71 mg of the title compound. MS (m / z): 533 (M + 1).

Example 55: 2-Chloro-1 '- [[1- (3-chloro-2-pyridyl) -3-methyl-pyrazol-4-yl] methyl] -4,4-difluoro-spiro (L) - [5H-thieno [2,3-c] pyran-7,4'-piperidine]

To a solution of 2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] (0.2 g, Is added 1- (3-chloro-2-pyridyl) -3-methyl-pyrazole-4-carbaldehyde (0.19 g, 0.86 mmol) in 1,2-dichloroethane (5 mL) and a few drops of acetic acid. The reaction mixture is stirred at room temperature for 1 hour. 102 ΡΕ2501703

Sodium triacetoxyborohydride (0.315 g, 1.43 mmol) is added and stirred at room temperature for 14 hours. After completion, the reaction mixture is diluted with dichloromethane and washed with saturated sodium bicarbonate solution and brine. The organic phase is dried over sodium sulfate and concentrated in vacuo. The crude mixture is purified by HPLC to give 0.195 g (56%) of 2-chloro-1 '- [[1- (3-chloro-2-pyridyl) -3-methyl-pyrazol-4-yl] yl] -4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine]. MS (m / z): 485 (M + 1). The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 485 (M + 1).

Example 56: 2-Chloro-1 '- [[1- (3-cyclopropyl-2-pyridyl) -3-methyl-pyrazol-4-yl] methyl] -4,4-difluoro-spiro (L) - [5H-thieno [2,3-c] pyran-7,4'-piperidine]

The title compound is prepared essentially as described in Example 55 using 2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c ] pyran] and 1- (3-cyclopropyl-2-pyridyl) -3-methyl-pyrazole-4-carbaldehyde as starting materials in 41% yield. MS (m / z): 491 (M + 1). 103 ΡΕ2501703

Example 57: 4 - [[2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-theno- [2,3-c] pyran-7,4'-piperidine] yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methyl] morpholin-3-one

A mixture of 2-chloro-4,4-difluoro-1 '- [(3-methyl-1H-pyrazol-4-yl) methyl] spiro [5H-thieno [2,3- c] pyran-7,4' piperidine] (0.20 g, 0.53 mmol), 4 - [(2-bromo-3-pyridyl) methyl] morpholin-3-one (0.21 g, 0.77 mmol), copper (I) (0.015 g, 0.078 mmol) and cesium carbonate (0.36 g, 1.10 mmol) in dimethylformamide (5 mL) is degassed by bubbling with argon for 15 minutes. (1R, 2R) -N, N'-dimethylcyclohexane-1,2-diamine (0.02 g, 0.14 mmol) is added while degassing occurs for 15 minutes and then the mixture is heated to 130øC for 16 hours. Upon completion, the reaction mixture is allowed to cool to room temperature and filtered through celite. The residue is washed with ethyl acetate (2 x 25 mL) and the filtrate is washed with water (30 mL). The aqueous layer was extracted with ethyl acetate (2 x 25 mL) and the combined organic extracts were dried over sodium sulfate and concentrated in vacuo. The crude mixture is purified by column chromatography on silica gel eluting with dichloromethane / methanol (96: 4) to give 0.09 g (30%) of the titled compound. MS (m / z): 564 (M + 1). 104 ΡΕ2501703

The compounds of Examples 58-59 are prepared essentially as described in Example 57 from the intermediates described in Preparations 49 and 50 respectively.

Example 60: [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1- (3-chloro-2-pyridyl) pyrazol-3-yl] methanol F.

F

Cl

N

Cl HO N. ' tf

105 ΡΕ2501703 1. 4 - ((21-Chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,71-thieno [2,3-c] pyran] -1 -yl) methyl) -1- (3-chloro-2-pyridyl) -1H-pyrazole-3-carboxylate To a solution of 2'-chloro-4 ', 4'-difluoro- dihydroespiro [piperidine-4,7'-thieno [2,3-c] pyran] (0.35 g, 1.1 mmol) and 1- (3-chloropyridin-2-yl) -4-formyl-pyrazole- 3-carboxylate (0.3 g, 1.0 mmol) in 1,2-dichloroethane (20 mL) is added N-methylmorpholine (0.33 g, 3.2 mmol) and molecular sieves (0.10 g ). The reaction mixture is stirred at room temperature for 1 h. Sodium triacetoxyborohydride (0.58 g, 2.7 mmol) is added and stirred at room temperature for 16 h. After completion, the reaction mixture is filtered through celite and partitioned between dichloromethane (15 mL) and water (15 mL). The aqueous phase is extracted with dichloromethane (3 x 30 mL) and the combined organic extracts are dried over sodium sulfate and concentrated in vacuo. The crude mixture is purified by column chromatography on silica gel eluting with dichloromethane / methanol (98: 2) to afford 0.4 g (67%) of 4 - ((2'-chloro-4 ', 4'- difluoro-4 ', 5'-dihydrospiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-yl) methyl) -1- (3-chloro-2-pyridyl) -1H -pyrazole-3-carboxylate. MS (m / z): 543 (M + 1). 2. [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1- ( To a solution of 4 - ((2'-chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-2-carboxylic acid 3-chloro-2'-pyridyl) pyrazol- 4,7'-thieno [2,3-c] pyran] -1-yl) methyl) -1- (3-chloro-2-pyridyl) -1H-pyrazole-3-carboxylate (0.4 g, 0.73 mmol) in tetrahydrofuran (5 mL) and ethanol (5 mL) is added lithium borohydride (1.80 mL, 2.0M solution in tetrahydrofuran, 3.68 mmol) at 0 ° C and stirred at room temperature for 16 h. After completion, the reaction mixture was quenched with water (10 mL) and extracted with ethyl acetate (2 x 25 mL). The combined organic extracts are dried over sodium sulfate and concentrated in vacuo. The crude mixture is purified by column chromatography on silica gel eluting with dichloromethane / methanol (97: 3) to give 0.14 g (39%) of the titled compound. MS (m / z): 500 (M + 1).

Example 61: [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1- (3-methyl-2-pyridyl) pyrazol-3-yl] methanol

The title compound is prepared essentially as described in Example 60 using 2'-107 ΡΕ2501703 hydrochloride chloro-4 ', 4'-difluoro-4', 5'-dihydrospiro [piperidine-4,7'-thieno [2 , 3-c] pyran] and 1- (3-methylpyridin-2-yl) -4-formyl-pyrazole-3-carboxylate as starting materials in 14% yield. MS (m / z): 481 (M + 1).

Example 62: [2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl ] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol

1. 2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3 methyl-pyrazol-1-yl] pyridine-3-carbaldehyde

A mixture of 2-chloro-4,4-difluoro-1 '- [(3-methyl-1H-pyrazol-4-yl) methyl] spiro [5H-thieno [2,3- c] pyran-7,4' (131.9 g, 57.7 mmol) and potassium carbonate (131 g, 947 mmol) in anhydrous toluene (340 mL) is degased at room temperature for 30 minutes. min. 2-Bromo-pyridine-3-carboxaxaldehyde (125 g, 677 mmol) and trans-N, N'-dimethylcyclohexane-1,2-diamine (21.3 mL, 135.4 mmol) are added sequentially. The contents are degassed for 30 min before heating to 105øC while shaking for 18 h. Analysis by LC / MS of the 108 ΡΕ2501703 reaction mixture reveals complete consumption of 2-chloro-4,4-difluoro-1 '- [(3-methyl-1H-pyrazol-4-yl) methyl] spiro [5H- 2,3-c] pyran-7,4'-piperidine]. The reaction mixture is cooled to room temperature, diluted with ethyl acetate (1.5 L), stirred and filtered through a pad of celite. The mother liquor is washed sequentially with 10% ammonium hydroxide (5 x 100 mL), water (3 x 100 mL), and brine. It is then dried (sodium sulfate), filtered and concentrated under reduced pressure. The residue is purified on silica gel using 50% ethyl acetate in hexane containing 1% triethylamine to provide 2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3- piperidin] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] pyridine-3-carbaldehyde as a solid (156 g, 72% yield) . MS (m / z): 479 (M + 1). 2. [2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyrazol-7, 4'-piperidine] -1'-yl) To a solution of 2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyrimidin- pyran-1-yl] pyridine-3-carbaldehyde (180 g, 377 mmol) in anhydrous dichloromethane (1.5 L) at 0 ° C is added sodium borohydride (7.2 g, 188.5 mmol) and anhydrous methanol (0.5 L) sequentially and the contents are allowed to reach room temperature while stirring is continued for 30 min. LC / MS analysis of the reaction mixture reveals conclusion. The volatiles are removed under reduced pressure and the residue obtained is partitioned between dichloromethane (2 L) and water (300 ml). The layers are separated and the organic layer is sequentially washed with 1N aqueous sodium hydroxide solution (300 mL), water (3 x 300 mL), brine, dried (sodium sulfate), filtered and concentrated under reduced pressure. The residue is purified on silica gel using 50-55% ethyl acetate in hexane containing 1% triethylamine to give [2- [4 - [(2-chloro-4,4-difluoro-spiro [5H- thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol as a solid (167 g, 92% yield). MS (m / z): 481 (M + 1).

[2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol

[2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3- methyl-pyrazol-1-yl] -3-pyridyl] methanol (157.4 g, 328 mmol) in ethyl acetate (1.5 L) is heated at 80 ° C until a clear solution is obtained. To the hot solution containing free base is 110 ΡΕ2501703 slowly added 5-6N HCl in isopropanol (65.6 mL, 328 mmol) and the contents are stirred vigorously while allowing the mixture to reach room temperature for 2 h. Precipitation is observed along the addition of HCl. The resulting white solid is filtered, washed with diethyl ether (3 x 1 L), dried under vacuum at 50 ° C for 3 days to obtain [2- [4 - [(2-chloro-4,4-difluoro -spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol as a solid (165 g, 97% yield). MS (m / z): 481 (M + 1).

Receptor Occupancy Compound Compound: 2 - [(2-Fluorophenyl) methyl] -3- (2-fluoro-spiro [4,5-dihydrothieno [2,3-c] pyran-7 , 4'-piperidine] -1'-yl) -N, N-dimethylpropanamide

1. tert-Butyl 3- (2-fluorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) propanoate 2-Fluorospiro [4.5 (2.7 g, 11.9 mmol) is dissolved in methanol (60 mL). Then, triethylamine (2.65 mL) and tert-butyl acrylate (3.55 mL, 23.76 mmol) are added and the mixture is heated to 65 øC for 5 h. The heating is removed and the reaction mixture is stirred at room temperature overnight. The solvent is evaporated and the crude product is purified by Isco normal phase chromatography using 1/1 ethyl acetate / hexane as eluent to afford 4.2 g of the desired compound as a colorless oil. MS (m / z): 356 (M + 1). 2 - [(2-Fluorophenyl) methyl] -3- (2-fluoro-spiro [4,5-dihydro-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl ) propanoate To a stirred solution of tert-butyl 3- (2-fluorospiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) propanoate tert-butyl (4.9 g, 13.78 mmol) in tetrahydrofuran (41 mL) under N 2 and cooled to -78 ° C, 1M lithium bis (trimethylsilyl) amide 35 mL, 41.45 mmol). The resulting mixture is stirred at that temperature for 3 hours. Then, 1,3-dimethyl-3,4,5,6-tetrahydro-2 (1H) -pyrimidinone (1.33 mL, 11.03 mmol) is added, and the resulting solution is stirred at the same temperature for 30 min. To the resulting mixture, 2-fluorobenzyl bromide (2.33 mL, 19.3 mmol) in dry tetrahydrofuran (1 mL) is added and stirring is continued. The temperature is allowed to rise from -78 ° C to room temperature overnight. The crude reaction mixture is quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer is decanted, dried over magnesium sulfate, the solvent is evaporated and the crude product obtained is purified by standard Isco phase chromatography using ethyl acetate / hexane from 5/95 to 20/80 to afford 5- 06 g of the title compound obtained as a colorless oil. MS (m / z): 464 (M + 1). 3. 2 - [(2-Fluorophenyl) methyl] -3- (2-fluoroespiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1 ' -yl) propanoic acid

A mixture of 2 - [(2-fluorophenyl) methyl] -3- (2-fluoroespiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) propanoate (5.06 g, 10.91 mmol) and trifluoroacetic acid (26.20 mL, 218 mmol) is stirred at room temperature overnight. The solvent is evaporated to dryness and the crude product is used without further purification. 4 - [(2-Fluorophenyl) methyl] -3- (2-fluoro-spiro [4,5-dihydro-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl ) - N, N -dimethylpropanamide

2 - [(2-Fluoro-phenyl) methyl] -3- (2-fluoro-spiro [4,5-dihydrothieno [2,3-c] pyran-7,4 '-piperidine] -1' -yl) propanoic acid (5.68 g, 10.89 mmol) is dissolved in dichloromethane (218 mL), then triethylamine (12.14 mL, 87.13 mmol), dimethylamine hydrochloride (1.80 g, 21.78 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (4.18 g, 21.78 mmol), and hydrate of 1-113 Ρ TE2501703 hydroxybenzotriazole (3.34 g, 21.78 mmol) are subsequently added to the solution at 0 ° C. The mixture is stirred at room temperature for 5 hours. The reaction mixture is treated with saturated aqueous sodium bicarbonate solution and extracted with dichloromethane (3 x 20 mL). The combined organic layers are dried over magnesium sulfate and the solvent is evaporated under reduced pressure. The crude product is purified by Isco normal phase chromatography using dichloromethane / 2N ammonium in methanol from 100/0 to 90/10 as eluant to give 4.0 g (84.5%) of the title compound. MS (m / z): 435 (M + 1). 5. (L) -2 - [(2-Fluorophenyl) methyl] -3- (2-fluorospiro [4,5-dihydrothieno [2,3-c] pyran-7,41-piperidine] The enantiomeric resolution of 2 - [(2-fluorophenyl) methyl] -3- (2-fluoro-spiro [4,5-dihydrothieno [2,3-c] ] pyran-7,4'-piperidine] -1'-yl) -17,7-dimethylpropanamide (2.3 g, 5.29 mmol) is run with a Chiralpak AD® column (Chiral Technologies, Inc. , West Chester, Pennsylvania, USA) using hexane / dimethylethylamine 0.2% in ethanol 9/1. The desired compound is obtained in 36% yield as the first eluting enantiomer. The tartrate salt is essentially prepared as described in Example 1. MS (m / z): 435 (M + 1). 114 ΡΕ2501703

Powder X-ray Diffraction The XRD pattern of the crystalline solid is obtained on a Bruker D4 Endeavor powder X-ray diffractometer, equipped with a CuKa λ = 1.54060 Âμ source) and a Vantec detector, which operates at 35 kV and 50 mA. The sample is scanned between 4 and 40ø in 20 with a pitch size of 0.0087ø in 2θ and a scan rate of 0.5 sec / in and with 0.6 mm of divergence, anti-dispersion set at 5.28, and 9.5 mm detector slits. The dry powder is packed into a quartz sample holder and a smooth surface is obtained using a glass slide. The diffraction patterns of the crystalline form are collected at ambient temperature and relative humidity. The bottom was removed before peak selection. It is well known in the art of crystallography that, for any given crystalline form, the relative intensities of the diffraction peaks may vary due to the preferred orientation resulting from factors such as crystalline morphology and habit. When the effects of the preferred orientation are present, peak intensities are altered, but the characteristic positions of the polymorph peaks do not change. See, e.g., The United States Pharmacopeia # 23, National Formulary # 18, pages 1843-1844, 1995. Furthermore, it is also well known in the art of crystallography that for any given crystalline form the angular positions of the peak may vary slightly. For example, peak positions may deviate due to a variation in the temperature or humidity at which a sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variability of ± 0.1 ° in 20 will take these potential variations into account without impeding the unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form may be made on the basis of any single combination of distinct peaks (in units of ° 2Θ), typically the most prominent peaks.

Thus, a crystalline sample prepared from the free base of (2- {4 - [(2'-chloro-4 ', 4'-difluoro-4', 5'-dihydro-1H-spiro [piperidine- 7'-thieno [2,3-c] pyran] -1-yl) methyl] -3-methyl-1H-pyrazol-1-yl} pyridin-3-yl) methanol is characterized by an XRD standard using CuKa radiation as having diffraction peaks (2-theta values) as described in Table 1 below. The single crystalline form can be confirmed with a subset of dintintos peaks from this complete diffraction pattern. Thus, in one embodiment the present invention provides a crystalline form of (2- {4 - [(2'-chloro-4 ', 4'-difluoro-4', 5'-dihydro-1H-spiro [ piperidin-4,7'-thieno [2,3-c] pyran] -1-yl) methyl] -3-methyl-1H-pyrazol-1-yl} pyridin-3-yl) methanol characterized by an XRD standard using CuKa radiation having diffraction peaks at 11.1 in combination with one or more of the peaks selected from the group consisting of 5.5, 13.5, 17.8, and 22.3 ± 0.1 ° in 2θ. In another embodiment, the present invention provides a crystalline form of (2- {4 - [(2'-chloro-4 ', 4'-difluoro-4', 5'-dihydro-1H-spiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-yl) methyl] -3-methyl-1H-pyrazole-1-yl] methyl] pyridin-3-yl} methanol XRD standard using CuKa radiation having diffraction peaks at 5.5, 11.1, 13.5, 17.8, and 22.3 ± 0.1 ° at 2Θ.

Table 1: X-ray diffraction peaks of the free base powder of (2- {4 - [(2,2-chloro-4,4'-difluoro-4 ', 5'-dihydro-1H-spiro [ piperidin-4,7'-thieno [2,3-c] pyran] -1-yl) methyl] -3-methyl-1H-pyrazol-1-yl} pyridin-3-yl) methanol

Peak Angle (° 2 theta) Intensity% 1 5, 5 53.2 2 11.1 100 3 12.3 9.9 4 13.5 12 5 15.4 5.1 6 16.6 6 4.8 7 17, 0 5.5 8 17.8 12.1 9 19.3 3 10 10 19.5 5.5 11 19.7 75.6 12 21.0 11.1 13 22.3 25.834 23.3 10.3 15 23.9 9.5 16 26.5 5.2 17 27.0 7.1 117 ΡΕ2501703

Likewise, a crystalline sample prepared from the HCI salt of (2- {4 - [(2'-chloro-4 ', 4'-difluoro-4', 5'-dihydro-1H-spiro [piperidine-4 , 7'-thieno [2,3-c] pyran] -1-yl) methyl] -3-methyl-1H-pyrazol-1-yl} pyridin-3-yl) methanol is characterized by a standard XRD using CuKa radiation as having diffraction peaks (2-theta values) as described in Table 2 below. The single crystalline form can be confirmed with a subset of distinct peaks from the complete diffraction pattern. Thus, in one embodiment the present invention provides a crystalline form of (2- {4 - [(2'-chloro-4 ', 4'-difluoro-4', 4'-dihydro- spiro [piperidine-4,7'-thieno [2,3-c] pyran-1-yl) methyl] -3-methyl-1H-pyrazol-1-yl} pyridin-3-yl) methanol an XRD pattern using CuKa radiation having diffraction peaks at 16.2 in combination with one or more of the peaks selected from the group consisting of 10.8, 12.1 and 21.1 ± 0.1 ° in 2θ. In another embodiment, the present invention provides a crystalline form of (2- {4 - [(2'-chloro-4 ', 4'-difluoro-4', 5'-dihydro-1H-spiro [piperidine-4,7'-thieno [2,3-c] pyran] -1-yl) methyl] -3-methyl-1H-pyrazol-1-yl} pyridin-3-yl) methanol characterized by a standard XRD using CuKa radiation having diffraction peaks at 10.8, 12.1, 16.2, and 21.2 ° ± 0.1 ° in 2θ.

Table 2: X-ray powder diffraction peaks of (2- {4 - [(2'-chloro-4 ', 4'-difluoro-4', 5'-dihydro-1H-spiro [ piperidin-4,7'-thieno [2,3-c] pyran] -1-yl) methyl] -3-methyl-1H-pyrazol-1-yl} pyridin-3-yl) methanol 118 ΡΕ2501703

Peak Angle (° 2 theta) Intensity% 1 10.8 70.5 2 12.1 41.4 3 16.2 80.1 4 18.7 60.8 5 19.5 38.66 20.649, 5 7 21.1 64, 9 8 22.3 33, 19 23, 89, 6 10 23.5 100 11 24.0 43.8

Data from the literature (Przydzial and Heisler, 2008, supra; Reinscheid, 2006, supra) and data generated from non-clinical animal studies support a role for nociceptin antagonists in the treatment of depression, obesity and eating disorders, and migraine. It is contemplated that nociceptin receptor antagonists are effective in rodent depression models either alone or in combination with selective serotonergic reuptake inhibitor (SSRI) or tricyclic antidepressants, inhibition inhibitors of rodent hyperphagia of weight loss following a previous weight loss, and in models for migraine. In addition, studies conducted in nociceptin receptor knockout mice have demonstrated that the action of nociceptin antagonists on the forced swimming test (measurement of antidepressant activity) and induced feeding after fasting (antiobesity activity) depends on the genotype , supporting a specific mechanism of nociceptin antagonistic action in these animal models. Since the disorders described above represent common co-morbid conditions, a nociceptin receptor antagonist may be particularly effective in these specific patient populations, such as patients with profound depression disorder, overeating disorder, overweight, obesity, and obesity with co-morbid clinical mood disorders.

To further the characterization of the present compounds, representative compounds are tested in the following in vitro and in vivo assays:

In Vitro Receptor Binding

Radioligand binding assays are commonly used to determine the affinity (Κ ±) or potency of a compound to bind to a particular receptor or target protein. A filtration based [3 H] -ORO / nociceptin receptor binding assay is developed based on prior assay formats (Ardati A, Henningsen RA, Higelin J, Reinscheid RK, Civelli 0, Monsma FJ Jr. Mol Pharmacol (May, 51 (5): 816-24.) With minor modifications. [3 H] -OFQ / nociceptin binding assays are performed in 96 well deep well plates. Competition studies [3H] OFQ (final concentration of the 0.2 nM assay) are carried out with 5-10æg of 120 ΡΕ2501703

membrane protein (isolated from Chinese hamster ovary cells (CHO cells) expressing cloned human ORL1 receptors) in a final assay volume of 0.5 ml of buffer containing 20 mM HEPES, pH 7.4, 5 mM MgCl 2 , 1 mM EGTA, 100 mM NaCl, 0.1% bovine serum albumin. The samples are incubated for 60 min at room temperature, which is considered to be optimal for competition assays. The assays are terminated by filtration through glass fiber filters (Wallac A filter mat) [pretreated with 0.3% polyethyleneimine (Sigma) for 1 h] in a Tometc cell collector, and the filters are washed three times with 5 mL of ice cold 50 mM Tris-HCl, pH 7.4. The filter mats are then dried and soaked in Meltilex A scintillant and the radioactivity counted in a Wallac Microbeta Scintillation Counter. Specific binding is determined by displacement with unlabeled 100 nM nociceptin. The curves are plotted as the percentage of specific binding and the IC 50 values are determined using a sigmoidal dose response curve with variable slope. The K + values are calculated from the IC50 by the equation of Cheng and Prusoff (Cheng, YC, and Prusoff, WH, Biochem., Pharmacol, 22, 3099-3108 (1973)) where Ki = IC 50 x (1 + D x Kd-1) &quot; 1.

Likewise, Ki for mu, kappa and delta opioids, serotonin, dopamine, adrenergic, mucarinin, and histamine receptors, as well as binding to the norepinephrine transporter, sodium channel, chloride channel, and calcium channel can be determined using membranes expressing the desired channels of the carrier / transporter and molecules that compete with the corresponding corresponding radioligands.

The exemplified compounds are tested essentially as described above and are considered to have high affinity for the ORL-1 Receptor. K ± for the ORL-1 Receptor for the exemplified compounds were shown to be less than 2 nM, whereas the K 2 for other tested recipients / transporters / channels were shown to be significantly higher. The compounds of Examples 62, 23, and 53 are tested essentially as described above and shown to have affinities as shown in Table 3 below.

Table 3. Selectivity data

Ki (nM) Example 62 Example 23 Example 53 ORL-1 0.18 0.79 0.45 Opioid Mu &gt; 451 &gt; 451 &gt; 451 Opioid & > 4780 5-HT1a> 3180 5-HT1A> 3180 5-HT1b> 3580 ND> 3580 5-HT1 ND ND ND 5-HT1d> 8550 ND> 8550 5-HT1e> 5370 ND> 5370 5-HTif> 8520 ND> 8520 5-HT2a> 5000 ND> 5000 5-HT2b 693 ND 220 122 ΡΕ2501703 (continued)

Ki (nM) Example 62 Example 23 Example 53 5-HT2c> 5650 ND> 5650 5-HT4> 4120 ND> 4120 5-HTs> 9090 ND> 90 90 5-HT6 ND ND> 5830 5 -HTv> 3980 ND> 3980 Dopamine Di> 5000 ND> 5000 Dopamine D2> 5000 ND> 5000 Adenosine A3> 5000 ND> 5000 Adrenergic Alfai> 5000 ND> 5000 Adrenergic Alpha 2> 5000 ND> 5000 Adrenergic Betai> 5000 ND> 5000 Adrenergic Beta2> 5000 ND> 5000 Histamine Hl> 5000 ND> 5000 Muscarinic M2> 500 ND> 5000 Muscarinic M3> 5000 ND> 5000 Carrier </ b> 500 ND &gt; 500 ND &gt; 500 ND &gt; 500 ND &gt; 500 ND &gt; 500 ND Channel> 500 ND> 500 ND Channel> 500 ND> 500 ND Channel local diltiazem)> 500 ND> 500 Ca + 2 channel (verapamil site)> 500 ND> 5000 ND = not determined

Accordingly, physiologically relevant doses of the compounds of the invention are not expected to interact 123 ΡΕ2501703 substantially with these sites in vivo, and thus it is expected to prevent undesired effects associated with such activity.

Functional In Vitro Functional Blockade of GTPy- [35 S] -Gonation of G-Protein Activation Mediated by Agonist

G-protein coupled receptor agonist-mediated stimulation results in the activation of membrane-associated heterotrimer-GaPY protein complexes and represents the first step in the transduction of extracellular signals for intracellular pathway modification. The first step in the activation of receptor-mediated heterotrimer GaPY proteins is the exchange of guanosine diphosphate bound to Ga (GDP) subunit by guanosine triphosphate (GTP). GTP binding to the Ga subunit causes dissociation of the heterotrimer subunits, GP and Gy, resulting in the modulation of several intracellular signaling cascades. Measurement of receptor-mediated G-protein activation can be measured using the non-hydrolyzable radiolabeled analog of GTP, GTP-γ [35 S]. Using this methodology, antagonist (Kb) affinity is measured on membranes expressing cloned human 0RL1 / nociceptin receptors using a GTP-γ- [35 S] binding assay according to protocols previously described with minor modifications (DeLapp et al. , J Pharmacol Exp Ther. 1999 May; 289 (2): 946-55; Ozaki et al., Eur J Pharmacol 2000 Aug 18; 402 (1-2): 45-53). The assays are conducted in a 200-μl volume with the following buffer composition: 100 mM NaCl, 20 mM HEPES, 5 mM MgCl2, 1 mM EDTA, 0.1% BSA at 124 ΡΕ2501703, 3 GDP GDP, [35 S] GTPγS 0.5 nM. The ORL1 receptor membrane suspension is added at a concentration of 20æg protein per well and receptor stimulation is achieved using 300 nM nociceptin / OFQ. Wheat germ agglutinin-coated SPA beads (Amersham, Arlington Hts., IL) are added at 1 mg per well to detect membrane bound [35 S] GTPyS. The plates are sealed and incubated for 2 h at room temperature. The plates are then placed at 4 ° C overnight to allow the SPA beads to stand and then counted on a Wallac Microbeta. The [35 S] GTPγS specific binding is determined to be the difference in CPM observed in the absence and presence of unlabeled 10 μM GTPγS. Data are shown graphically as the percent binding of [35S] GTPyS specific. Curves are plotted as the percentage of specific binding and IC 50 values are determined using the curve of

sigmoidal dose response with variable slope. The affinity of the antagonist (¾) is estimated according to DeLapp et al., 1999 using a modification of the equation of Cheng and Prusoff (1973) where Kb = IC 50 x (1 + D x EC 50 1) 1.

The exemplified compounds are tested essentially as described above and have been shown to be potent antagonists of the ORL-1 receptor. Kb for the ORL-1 Receptor for the exemplified compounds was shown to be less than 6 nM. The compounds of Examples 1, 23, and 53 are tested essentially as described above and shown to have Kb for the ORL-1 Receptor of 0.20, 1.52, and 0.62 nM, respectively. 125 ΡΕ2501703

Receptor Occupancy In Vivo Receptor occupancy (RO) using LC / MS / MS was established as a way to measure the central target impairment of the putative ORL-1 antagonist in vivo. The nociceptin / ORL1 (RO) receptor occupancy is measured in the hypothalamus, a structure containing a high density of nociceptin / ORL1 binding sites found in the blood-brain barrier, using a novel nociceptin / ORL1 antagonist protected RO marker, 2 - [(2-fluorophenyl) methyl] -3- (2-fluoroespiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) -N, N-dimethylpropanamide, (RO Marker). These measurements are made without the need for a radiolabeled label as previously published for other modified receptors (Chernet E, Martin LJ, Li D, Need AB, Barth VN, Rash KS, Phebus LA. In preclinical distribution, in vivo receptor occupancy studies: dopamine D2, serotonin 2A and NK-1 receptors as examples.Life Sei. 78 (4): 340-6, 2005.).

A positive correlation was established between central nociceptin / ORL1 RO and efficacy in modulating feeding behavior and forced swimming test in rodents. RO nociceptin / central ORL1 is measured 6 or 24 hours after oral administration of the test compound to rats. Male Sprague-Dawley rats (Harlan Sprague-Dawley, Indianapolis, IN) are treated orally with a test compound, or vehicle (Captisol 20%, 25 mM phosphate buffer, pH 2.0). 6 or 24 hours following the administration of the test compound / vehicle, a dose of 3æg / kg RO Marker is administered intravenously to all animals. It is at the time of administration of the RO Marker that RO is considered for measurement. Forty minutes after administration of the RO Marker, the rats are sacrificed by cervical dislocation and the hypothalamus is removed. The RO Marker level is measured on each tissue sample. The reference standard of the centrally active (-) - cis-1-methyl-7 - [[4- (2,6-dichlorophenyl) piperidin-1-yl] methyl] -6,7,8,9- tetrahydro-5H-benzocycloheptan-5-ol (SB612111, see Magdalena and Heisler, supra), a potent selective nociceptin / ORL1 receptor antagonist, is used as a positive control to establish the level of RO-associated RO Marker nociceptin / 100% ORL1. SB612111 is administered intravenously at a dose of 30 mg / kg 1 h before the RO Marker (a dose which results in approximately 100% RO of hypothalamic nociceptin / ORL1 receptors).

Hypothalamic samples are homogenized in 4 volumes (w / v) acetonitrile containing 0.1% formic acid and centrifuged at 14,000 RPM for 16 min. The supernatants are collected and diluted to a final volume of 0.3 mL with sterile water. Measurement of the RO Marker is performed using Agilent Model 1200 HPLC (Agilent Technologies, Paio Alto, CA) and an API mass spectrometer 127 ΡΕ2501703 4000. Chromatographic separation uses a 2.1 x 50 mm C18 column (Agilent part number 971700 -907) and a mobile phase consisting of 38% acetonitrile in water with an overall content of 0.1% formic acid. Detection of the RO Marker is achieved by monitoring the precursor to ion transition of the product at a mass to charge (m / z) ratio of 435 to 239, at quantified levels compared to standards prepared in homogenates of brain tissue. The zero percentage of RO is calculated as the RO Marker level in the hypothalamus of animals pretreated with vehicle, which represents the sum of non-specific and specific binding (all available receptors for the marker). The lowest level of RO Marker in animals pretreated with the very high intravenous dose of SB612111, the positive control group, represents non-specific binding and is assigned the 100% occupancy value (no available receptor for the marker). The level of the RO Marker found in the hypothalamus from the group treated with the test compound is linearly interpolated between these two points to calculate RO for the test compound.

The exemplified compounds are tested essentially as described above and have been shown to have high receptor uptake in ORL-1 Receptor. The occupations of the receptor for the exemplified compounds were found to be between about 40 to about 130% at 3 mg / kg dose after 6 h or between about 20 and about 96% RO at 3 mg / kg dose after 24 h . The occupations of the receptor 128 ΡΕ2501703 for the compounds of Examples 62, 23 and 53 are tested essentially as described above for dose of 3 mg / kg after 6 h and were shown to have RO 104, 80 and 83%, respectively. As such, the compounds of the present invention are expected to have bioavailability and CNS penetration favorable to the target ORL-1 Receptors.

Activity of the hERG channel The blockage of K + channel conductance in the heart is associated with cardiotoxicity in the form of QT wave extension. The affinity (K +) of the nociceptin receptor antagonists exemplified for human K + ERG (hERG) channel is determined in cloned hERG-expressing HEK293 cells using hERG channel antagonist radioligand [3 H] astemizole (2 nM final assay concentration) according to well-known procedures (see, for example, Finlayson K, et al. (Eur J Pharmacol 412 (3): 203-121, 2001). [3 H] Astemizole binding assays are carried out at the contracted research company Cerep Paris France) according to standard procedures.

Examples 62, 23, and 53 are essentially assayed as described above and have been shown to have reduced activity, with Ki of 6.08, 1.21, and 8.6 μΜ respectively. Comparison of plasma concentration in vivo required to produce 80 percent RO (ECsoSo) of nociceptin / ORL1 receptors in the CNS for the compound of example 62 is approximately 41 nM. As such, there is a large separation between the concentrations required to produce occupancy of the physiologically relevant in vivo receptor of the nociceptin / ORL1 receptors and the concentrations required for K + -hERG channel activity. Accordingly, physiologically relevant doses of the compounds of the invention are not expected to interact substantially with hERG sites in vivo, and thus are expected to have no substantial effect on the QT prolongation.

Mouse-Formed Test (mFST) mFST is an established in vivo assay for anti-depressant activity (Li et al., J Pharmacol Exp Ther. 319 (1): 254-9, 2006). Mice treated with known clinically effective anti-depressants (selective serotonin reuptake inhibitors and / or tricyclic antidepressants) exhibit the behavior of short-term immobility after being placed in a water tank, a behavior associated with despair. MFST was used to assess potential anti-depressant activity of novel nociceptin / ORL1 antagonists according to previously published methods (Li et al., J Pharmacol Exp Ther. 319 (1): 254-9, 2006). Briefly, male NIH-Swiss mice (Harlan Sprague-Dawley, Indianapolis, IN) weighing between 25-30 g. The group living animals are removed from the nursery to the test area in their own cages and allowed to adapt to the new medium for at least 1 hour before testing. Alternatively, male wild type 129S6 and 130 ΡΕ2501703 knockout mice relative to ORL1 / nociceptin receptors were used to confirm that the dependence on the response to the compound was dependent on the ORL1 / no-ciceptin receptor. All compounds are prepared in 20% Captisol, 25 mM phosphate buffer, pH 2.0 on the day of use. The mice are placed in a cylinder (diameter: 10 cm; height: 25 cm) filled with 6 cm of water (22-25 ° C) for 6 min. The duration of immobility during the last 4 min of the 6-min period of the test was noted. A mouse is recorded as immobile when it floats without movement or just making the movements necessary to keep the head out of the water.

Representative compounds are tested essentially as described above and have been shown to significantly reduce the immobilization times of wild type mice. The compounds of Examples 62, 23, and 53 are essentially assayed as described above and have been shown to have Π"β3 of 24.9, 9.5, and 12.9 mg / kg respectively, with maximum effects of 45%, 58%, and 54% reductions in immobilization time, respectively. Accordingly, the compounds of the present invention are expected to have anti-depressant activity in vivo.

In addition, the compounds of the present invention may be used in combination with other known anti-depressants to produce improved efficacy. The compound of Example 62 is tested essentially as 131 ΡΕ2501703 described above in combination with 10 mg / kg of fluoxetine and has been shown to significantly reduce the immobilization time over both fluoxetine and the compound of Example 62 alone. See Table 4 below.

Table 4 Forced Swim Testing of Mice - Treatment

Combined with fluoxetine

(N = 8) Vehicle 174 16 3 mg / kg of Ex. 62 163 17 10 mg / kg of Ex. 62 129 19 10 mg / kg of fluoxetine 168 14 3 mg / kg of Ex. 62 + 10 mg / kg of fluoxetine 97 * 19 10 mg / kg of Ex. 62 + 10 mg / kg of fluoxetine 85 15 15 mg / kg Imiprimin (positive anti-depressant control) 96 * 20 * Statistically significant reduction relative to the vehicle, Ex 62 alone, and fluoxetine alone.

In addition, these effects of reduced immobilization times are not observed when ORL-1 knockout mice, a strain of mice created so as to lack ORL-1 receptor, are used, demonstrating that in effect the effect is mediated by the ORL-1 receptor. The compound of Example 62 is tested essentially as described above using wild-type mice in one of the study groups and ORL-1 knockout mice in the second study group, and was shown to significantly reduce the time of immobilization in the wild-type mice but did not show effect on the knockout mice. A norepinephrine reuptake inhibitor, imipyrimine, is tested as a positive control and has been shown to reduce immobilization times in either wild type or knockout mice to an equivalent degree, demonstrating that the behavioral effect mediated by the norepinephrine reuptake mechanism is intact in the mouse knockout strain.

Blockade of Induced Hyperfagia after Fasting in Mice. Blocking of induced hyperphagia after fasting in rodents is an accepted model for hyperphagic eating disorders. (Hollopeter G, Erickson JC, Seeley RJ, Marsh DJ, Palmiter RD, Response of neuropeptide Y-deficient mice to feeding effectors, Regul Pept 1998 Sep 25; 75-76: 383-9.) All experiments are performed on mice 12-week-old male naïve naïve mice and ORL knockout mice maintained in a 129S6 congenital environment. The 133 ΡΕ2501703 mice are housed individually at least 3 days before the start of the tests to eliminate any stress effects due to the change of accommodation from group to individual housing. Three mice / genotype are randomly assigned to each treatment group on the day of the test. Measurements of pre-fast body weight are made, and the food is then removed from the cages overnight. Mice are fasted for approximately 15 h. The next morning, the mice are given one of three doses of the drug or vehicle orally by gastric tube 30 minutes before gaining access to the food. Drugs are dissolved in 20% Captisol dissolved in 25 mM phosphate buffer, pH 2.0. Body weight measurements are performed immediately prior to treatment by the drug or 24 hours after access to food is restored. It is worth noting that all mice regardless of genotype lose ~ 5-10% of body weight following overnight fasting. Measurements of food intake are recorded following food access, as indicated, by the weight of the food remaining after 1 h. It should be noted that the measurement of food intake is measured during the light phase, a period during which the mice are typically resting and are not normally eating. Following the initial test, the mice rested for 1 week with access to unlimited feed. Following the week of rest, the mice are retested according to the Latin square protocol presented in Table 5. 134 ΡΕ2501703

Table 5. A Latin square protocol is used to determine the dose response curves of the anorectic action of the novel ORL antagonists in wild-type mice and ORL knockout mice.

Treatment Period Treatment Group A B C D 1 Vehicle A c B 2 A B Vehicle C 3 B C A Vehicle 4 C Vehicle B A

Representative compounds are tested essentially as described above and have been shown to significantly reduce induced hyperphagia after fasting in mice. Examples 62, 23, and 53 are essentially assayed as described above and have been shown to substantially block post-fasting induced hyperphagia. The effect was not observed in the strain of ORL-1 knockout mice, demonstrating that the effect is mediated through the ORL-1 receptor. The 5-HT2o mCPP agonist is used as a positive control and has been shown to significantly reduce fasting induced hyperphagia in both wild-type mice and the strain of ORL-1 knockout mice. As such, the compounds of the present invention are expected to be useful in the treatment of overweight and / or obesity and / or for weight maintenance, such as over-eating treatment. 135 ΡΕ2501703

Durai Rat Plasma Protein Stapling (PPE) Model - Oral Dosing Protocol

All test compounds are prepared in a vehicle solution containing 20% Captisol in 25 mM phosphate buffer (pH 2.0). The positive control compound, sumatriptan, is dissolved in saline. To the male Sprague-Dawley rats of Harlan Laboratories (250 to 350 g), which were fasted overnight, doses of the test compound, sumatriptan or vehicle are given by oral gastric probe (2 ml / kg) . Fifty min after the dose the rats are anesthetized with Nembutal (60 mg / kg, ip) and placed on a stereotactic grate with the incisor bar adjusted to -2.5 mm. Following a longitudinal incision in the midline of the scalp, 2 pairs of bilateral holes were drilled through the skull (3.2 mm later, 1.8 and 3.8 mm laterally, all coordinates referenced to bregma). Pairs of Rhodes Medical Systems Inc, isolated except for the ends, are lowered through the holes in both hemispheres to a depth of 9.2 mm below the hard.

A fluoroscein isothiocyanate (FITC) (FITC-BSA) dye-labeled bovine serum albumin (BSA) solution (20 mg / kg, iv) is injected into the femoral vein 2 minutes prior to electrical stimulation of the trigeminal ganglion to function as marker for the protein entrapment. Sixty minutes following dosing with test compound or vehicle 136 ΡΕ2501703, the left trigeminal ganglion is electrically stimulated for 5 minutes at a current intensity of 1.0 mA (5 Hz, 5 minutes duration).

Five minutes after stimulation, the rats are killed by exsanguination with 40 mL of saline solution which also removes residual FITC / BSA by rinsing the blood vessels. The top of the skull is removed to collect the dural membranes. The membrane samples are removed from both hemispheres, rinsed with water, and scattered horizontally on microscope slides. The lamellae are dried for 15 minutes in a lamellar heater and covered with a coverslip with 70% glycerol / water solution.

A fluorescence microscope equipped with a network monochromator and a spectrophotometer is used to quantify the amount of FITC-BSA dye in each dura sample. The microscope is equipped with a monitored platform interfaced with a personal computer. This facilitates the movement of the computer controlled platform, with fluorescence measurements at 25 points (500 ym steps) on each sample duration. The entrapment induced by electrical stimulation of the trigeminal ganglion is an ipsilateral effect (which occurs only on the side of the dura where the trigeminal ganglion is stimulated). This allows the other (unstimulated) half of the hard to be used as a control. The ratio of the entrapment (which is the 137 ΡΕ2501703 ratio of the amount of entrapment in the hard of the stimulated side compared to the non-stimulated side) is calculated. Animals given vehicle-only doses or an ineffective dose of the test compound have a release ratio of approximately 2, while fully effective treatments result in a ratio of approximately 1.

Results are expressed as mean values with mean standard deviations (± SEM). All statistical evaluations are conducted using ANOVA followed by comparison with the control group by Dunnett's method. Statistical significance is assumed when p <0.05. Statistical analyzes are performed using JMP statistical analysis software (SAS Research Institute, version 6.0.2). The compound of Example 62 is tested essentially as described above and has been shown to effectively block the dosing in a dose-dependent manner. (See Table 5 below.) As a result, the compounds of the present invention are expected to be useful in the treatment of migraine.

Table 5. Proliferation of plasma protein (PPE) lasts in rats for the compound of Example 62. Vehicle. 138 ΡΕ2501703

Treatment (po) Spreading ratio (mean) s.e. Ex. 62 (1.0 mg / kg) 1.44 * 0, 08 3 Ex. 62 (0.1 mg / kg) 1.83 10.0 mg / kg) 1.20 * 0.03 3 Ex. 62 (30.0 mg / kg) 1.14 * 0.05 Sumatriptan (1.0 mg / kg) 1.06 * 0.01 3

Stability for formation of the reactive metabolite

Previous literature suggests a correlation between the formation of the reactive metabolite and clinical toxicities known as idiosyncratic drug reactions (IDRs), although no direct causal effect has been established. Assuming that reactive metabolites may play a role in clinical IDRs, it was proposed the minimization of the potential for oxidizing bioactivation as a means to improve the overall safety profile of compounds containing structural characteristics associated with such reactivity. (see Baillie, Thomas A., Approaches to the Assessment of Stable and Chemically Reactive Drug Metabolites in Early Clinical Trials, Chemical Research in Toxicology, vol 22 (2) 2009.) To this end, representative compounds of the present invention and related compounds are screened using a hepatic microsomal capture assay in rats using glutathione as an endogenous nucleophile to comprise the oxidant bioactivation potential of the Î »TE2501703 portion of the thienyl moiety. Of the compounds tested, those in which R2a and R2b are hydrogen have shown evidence of conjugated formation of glutathione suggestive of oxidation in the thienyl moiety. Of the compounds tested, those in which R2a and R2b are fluoro did not show conjugated formation of glutathione. (See Table 3 below.) The absence of conjugated glutathione formation for the gem-difluoro-containing molecules suggests that the gem-difluoro substituent reduces the inherent chemical propensity for bioactivation as tested in the assay.

Table 3. Conjugated formation of Glutathione in hepatic microsomal homogenate

Compound Conjugate formation of glutathione (L) 3-fluoro-2- [4 - [(2-fluoroespiro [4,5-dihydrothieno [2,3-c] pyran-7,4'-piperidine] - 1-yl) methyl] -3-methyl-pyrazol-1-yl] benzamide Yes (L) 3-Fluoro-2- [4 - [(2,3-difluoro-spiro [4,5-dihydro-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] benzamide Yes (Ex.14) (L) 2-Chloro- yl] methyl] spiro [4,5-dihydrothieno [2,3-c] pyran-3-yl] 7,4'-piperidine] Yes (Ex. 10) (L) 2,4,4-Trifluoro-1 '- [[1- [3- (methoxymethyl) -2-pyridyl] -3-methylpyrazole -4-yl] methyl] spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] No (Ex.18) (L) 3-fluoro-2- [3- 4 - [(2,4,4-trifluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] pyrazol-1-yl] benzamide No 140 ΡΕ2501703

While it is possible to administer compounds employed in the methods of this invention directly without any formulation, the compounds are usually administered in the form of pharmaceutical compositions comprising at least one compound of Formula I, or a pharmaceutically acceptable salt thereof, as an active ingredient and less a diluent carrier and / or pharmaceutically acceptable excipient. These compositions may be administered via a variety of routes including oral, intranasal, transdermal, subcutaneous, intravenous, intramuscular, and pulmonary. Such pharmaceutical compositions and processes for their preparation are well known in the art. See, e.g., Remington: The Science and Practice of Pharmacy (University of the Sciences in Philadelphia, ed., 21st ed., Lippincott Williams & Wilkins Co., 2005).

The compositions are preferably formulated in a unit dosage form, each dosage containing from about 0.1 to about 500 mg, more usually from about 1.0 to about 200 mg, for example from about 5 to 50 mg of the active ingredient. The term &quot; unit dosage form &quot; refers to suitable physically discrete units as unit dosages for human subjects and other mammals, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect in association with at least one carrier, diluent and / or excipient suitable pharmaceutically acceptable salt. 141 ΡΕ2501703

The compounds of Formula I are generally effective over a wide range of dosages. For example, daily dosages are usually within the range of about 0.01 to about 50 mg / kg, more usually between about 0.05 to 5.0 mg / kg, and for example between 0.1 and 1.0 mg / kg body weight. In some cases dosage levels below the lower limit of the above range may be more than adequate, while in other cases even larger doses may be employed without causing any detrimental side effect, and therefore the said dosage range is not intended to be in any way limiting the scope of the invention. It will be understood that the amount of the compound actually administered will be determined by a physician in the light of the relevant circumstances, including the condition being treated, the route of administration chosen, the actual compound or compounds administered, the patient's age, weight, and response individual, and the severity of the patient's symptoms.

Lisbon, November 20, 2013

Claims (7)

A compound of the formula: wherein R1 is fluoro or chloro; R 2a and R 2b are each hydrogen or are each fluoro; R 3 is hydrogen, methyl, hydroxymethyl, or (C 1 -C 3) alkoxy-methyl; R 4 is selected from the group consisting of fluoro, chloro, cyano, cyanomethyl, (C 1 -C 3) alkyl, cyclopropyl, hydroxymethyl, methoxy, cyclopropylmethoxy, aminocarbonylmethoxy, (C 1 -C 3) alkoxymethyl, cyclopropyloxymethyl, cyclopropylmethoxymethyl, hydroxy-1-methylethyl, aminocarbonyloxy-methyl, methylaminocarbonyloxymethyl, dimethylaminocarbonyl-xymethyl, aminocarbonyl, aminocarbonylmethyl, -CH 2 -NR 5 R 6, hydroxyimine, methoxyimine, morpholin-4-yl, morpholin-4-ylmethyl, Ar 1, -CH 2 Ar 1 tetrahydrofuran-2-yl, 3-oxomorpholin-4-ylmethyl, 2-oxopyrrolidin-1-ylmethyl, and 2-oxopiperidin-1-ylmethyl; 2 ΡΕ2501703 R5 is hydrogen, C1 -C3 alkyl, cyanomethyl, -C (O) CH3, or aminocarbonylmethyl; R1 is hydrogen or methyl; and Ar 1 is the moiety selected from the group consisting of imidizol-1-yl, imidizol-2-yl, 2-methylimidizol-1-yl, pyrazol-1-yl, 1,2,3-triazol-1-yl; 1,2,3-triazol-2-yl; 1,2,4-triazol-1-yl, isoxazol-3-yl, oxazol-5-yl, and 3-methyl-1,2,4-oxadiazol-5-yl; or a pharmaceutically acceptable salt thereof. The compound of Claim 1 wherein R1 is chloro, or a pharmaceutically acceptable salt thereof. The compound of Claim 1 or 2 wherein R2a and R2b are each fluoro, or a pharmaceutically acceptable salt thereof. The compound of Claim 1 wherein R1 is fluoro and R2a and R2b are each hydrogen, or a pharmaceutically acceptable salt thereof. The compound of any one of Claims 1 to 4 wherein R 3 is methyl, or a pharmaceutically acceptable salt thereof. The compound of any one of Claims 1 to 5 wherein R 4 is fluoro, hydroxymethyl, methoxy-methyl, or pyrazol-1-ylmethyl, or a pharmaceutically acceptable salt thereof. The compound of Claim 1 which is [2- [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] -pyran-7,4 '-piperidine] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol; 3-methyl-1- [3- (pyrazol-1-ylmethyl) -2-pyridyl] pyrazol-4-yl] methyl] spiro [5H-thieno [ 2,3-c] pyran-7,4'-piperidine]; or [4 - [(2-chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] pyran-7,4'-piperidine] -1'-yl) methyl] -1- (3- Fluoro-2-pyridyl) pyrazol-3-yl] methanol; or a pharmaceutically acceptable salt thereof. The compound of Claim 1 which is [2- [4 - [(2-Chloro-4,4-difluoro-spiro [5H-thieno [2,3-c] -pyran-7,4'- piperidinyl] -1'-yl) methyl] -3-methyl-pyrazol-1-yl] -3-pyridyl] methanol, or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising a compound according to any one of Claims 1 to 8, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient. The pharmaceutical composition of Claim 9 further comprising at least one additional therapeutic ingredient. The pharmaceutical composition of Claim 4 wherein the additional therapeutic ingredient is an SSRI antidepressant. A compound according to any one of Claims 1 to 8, or a pharmaceutically acceptable salt thereof, for use in A compound or acceptable, for use in the treatment of obesity or A compound or acceptable for use in the treatment of migraine. A compound or acceptable for use in the treatment of depression. therapy. a pharmaceutically acceptable salt thereof according to claim 12 in a human. a pharmaceutically acceptable salt thereof according to Claim 12, a pharmaceutically acceptable salt thereof. The reference list cited by the applicant is for the reader's convenience only. It is not part of the European patent document. While due care has been taken in compiling references, errors or omissions may not be excluded and the IEP declines any liability in this regard. 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